Confocal NEMA?
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** hi all, anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? is anyone aware of this issue ever having been discussed by NEMA? cheers kurt . Prof. Kurt I. Anderson Tumor Cell Migration Lab and Beatson Advanced Imaging Resource (BAIR) The Beatson Institute for Cancer Research Garscube Estate, Switchback Road, Glasgow G61 1BD • Direct Line +44 (0) 141 330 2864 • Fax +44 (0) 141 942 6521 [hidden email]<mailto:[hidden email]> A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Kurt, I'm not familiar with NEMA, but the general topic of testing/standardizing confocal sensitivity, resolution, and image quality is of great interest to me. If you search the confocal listserver archive, you'll find several postings about this. In theory, you could develop a standard test for confocal sensitivity, resolution, and image quality, but what should the test specimen be? Others have worked on this to some extent, and a few names that come to mind are Fred Brakenhoff, Claire Brown, Robert Zucker, and Mike Model. I recommend looking up their papers to see what sorts of test specimens have been proposed and tried in the past. Unfortunately, I don't think there is one single test specimen that can be used to determine all instrument parameters of interest (sensitivity, resolution - spatial, temporal, spectral-, and image quality, etc.). In my mind, an ideal test specimen for confocal fluorescence imaging would have the following properties: 1. Be cheap, readily available, and reproducible to a high degree 2. Able to absorb and emit light over broad range of wavelengths 3. Be portable and emit constant intensity (no photobleaching) perhaps even with a known number of photons under certain conditions 4. Does not saturate easily, and has a linear response to excitation light power 5. Has a known 3D structure of specific shapes/patterns/sizes with a high level of precision determined by some other imaging technique (electron microscopy for example). 6. Has a refractive index close to water or glass Again, I don't think such a sample exists, but if you can think of one, let me know! I think one of the other problems here is: how would you define and quantify things like "image quality"? I'm curious how NEMA does this for PET. Spatial resolution is a bit more straightforward to quantify, although here too there are different ways to define resolution and ways to measure it. Sensitivity measurements are complicated by the need to keep many other imaging parameters constant when examining day-to-day performance/sensitivity of one instrument, or comparing the performance of two different instruments - see Jim Pawley's "39 steps". Your question also brings to mind a quote from another very well-written article on the topic which I never forget: "This complex relationship between qualitative and quantitative content of fluorescence images also expresses itself in the way that commercial instruments are assessed. On the one hand it is unavoidable that users initially are strongly influenced by the visual quality of the images presented by the instrument. On the other hand the longer term scientific value of the instrument depends crucially both on the quality of the visual information and the effectiveness with which it can be reduced for quantitative analysis... The increased complexity of a fluorescent image (in terms of the number of dimensions that can be measured), and the variability of sample preparation, makes it less easy to assess quantitatively the quality of the image. This may seem a surprising, or at least somewhat bleak, assessment. But consider this: In the 20 years or more since introduction of laser scanning confocal microscopes it has not been feasible, from published data, to assess how these systems compare (in terms of absolute units associated with measurements). No one can assert with confidence that instrument A in use in 1990 has better or worse sensitivity than instrument B operating in 2006. There is, therefore, a paramount need for standardized test samples and procedures for their use." -taken from: Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization of fluorescence measurements from the instrument manufacturer's view standardization and quality assurance in fluorescence measurements ii. 2008. 6: p. 3-24. I find the second last statement above quite frustrating, especially from the point of view of someone who works in the industry. I can tell you that the experience of demoing a microscope system to a potential customer often goes something like this: The potential customer prepares a typical biological specimen that they are familiar with from experience and then images of this specimen are captured with the demo system. These images are then compared to a similar set of images that were produced on another competing instrument (maybe - sometimes comparison images are not available). Usually there are statements made to the effect of "yes/no, the images on this instrument look better/worse, therefore I should buy/decline instrument A/B". I think anyone with a good imagination can figure out that this is a very subjective way to compare instrument performance and that there are a lot of potential pitfalls involved that might bias the assessment one way or another. I'm not saying this isn't a good method to judge an microscope's general imaging performance. Indeed, you should always take a prospective microscope system out for a "test drive", but a more quantitative test that doesn't solely depend on the opinion of how "good" an image looks would be better for everyone in my opinion. And Kurt, supposing a good set of text specimens and protocols were developed - how could we all come to agree on them? Who would set the standards and govern them for microscopy? Surely the industry players would have much to say (or dispute!) about that. Again, as stated above, there is a paramount need in microscopy for standardized test samples and procedures. It's a cause I certainly would be willing to devote time to were there an organized and concerted effort to make it happen. Curious to hear other people's thoughts on this. John Oreopoulos Staff Scientist Spectral Applied Research Inc. A Division of Andor Technology Richmond Hill, Ontario Canada www.spectral.ca On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > > > hi all, > > anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. > > they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. > > see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. > > recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. > > among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. > > my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? > > is anyone aware of this issue ever having been discussed by NEMA? > > cheers > > kurt > > . > > > > > Prof. Kurt I. Anderson > Tumor Cell Migration Lab and > Beatson Advanced Imaging Resource (BAIR) > The Beatson Institute for Cancer Research > Garscube Estate, Switchback Road, Glasgow G61 1BD > > > > • Direct Line +44 (0) 141 330 2864 > • Fax +44 (0) 141 942 6521 > > [hidden email]<mailto:[hidden email]> > > > > A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 > > > > The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** There’s a commercially available product that claims to meet most of the requirements John lists, below. Has anyone used this slide? http://argolight.com/product-micro/ My Best, — Glenn Glenn Merrill-Skoloff Division of Hemostasis and Thrombosis Director, Intravital Microscopy Core 617-735-4040 (Office) 617-735-4007 (Lab) 617-735-4000 (Fax) From: John Oreopoulos <[hidden email]<mailto:[hidden email]>> Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]>> Date: Saturday, March 15, 2014 at 12:35 AM To: "[hidden email]<mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]>> Subject: Re: Confocal NEMA? ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Kurt, I'm not familiar with NEMA, but the general topic of testing/standardizing confocal sensitivity, resolution, and image quality is of great interest to me. If you search the confocal listserver archive, you'll find several postings about this. In theory, you could develop a standard test for confocal sensitivity, resolution, and image quality, but what should the test specimen be? Others have worked on this to some extent, and a few names that come to mind are Fred Brakenhoff, Claire Brown, Robert Zucker, and Mike Model. I recommend looking up their papers to see what sorts of test specimens have been proposed and tried in the past. Unfortunately, I don't think there is one single test specimen that can be used to determine all instrument parameters of interest (sensitivity, resolution - spatial, temporal, spectral-, and image quality, etc.). In my mind, an ideal test specimen for confocal fluorescence imaging would have the following properties: 1. Be cheap, readily available, and reproducible to a high degree 2. Able to absorb and emit light over broad range of wavelengths 3. Be portable and emit constant intensity (no photobleaching) perhaps even with a known number of photons under certain conditions 4. Does not saturate easily, and has a linear response to excitation light power 5. Has a known 3D structure of specific shapes/patterns/sizes with a high level of precision determined by some other imaging technique (electron microscopy for example). 6. Has a refractive index close to water or glass Again, I don't think such a sample exists, but if you can think of one, let me know! I think one of the other problems here is: how would you define and quantify things like "image quality"? I'm curious how NEMA does this for PET. Spatial resolution is a bit more straightforward to quantify, although here too there are different ways to define resolution and ways to measure it. Sensitivity measurements are complicated by the need to keep many other imaging parameters constant when examining day-to-day performance/sensitivity of one instrument, or comparing the performance of two different instruments - see Jim Pawley's "39 steps". Your question also brings to mind a quote from another very well-written article on the topic which I never forget: "This complex relationship between qualitative and quantitative content of fluorescence images also expresses itself in the way that commercial instruments are assessed. On the one hand it is unavoidable that users initially are strongly influenced by the visual quality of the images presented by the instrument. On the other hand the longer term scientific value of the instrument depends crucially both on the quality of the visual information and the effectiveness with which it can be reduced for quantitative analysis... The increased complexity of a fluorescent image (in terms of the number of dimensions that can be measured), and the variability of sample preparation, makes it less easy to assess quantitatively the quality of the image. This may seem a surprising, or at least somewhat bleak, assessment. But consider this: In the 20 years or more since introduction of laser scanning confocal microscopes it has not been feasible, from published data, to assess how these systems compare (in terms of absolute units associated with measurements). No one can assert with confidence that instrument A in use in 1990 has better or worse sensitivity than instrument B operating in 2006. There is, therefore, a paramount need for standardized test samples and procedures for their use." -taken from: Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization of fluorescence measurements from the instrument manufacturer's view standardization and quality assurance in fluorescence measurements ii. 2008. 6: p. 3-24. I find the second last statement above quite frustrating, especially from the point of view of someone who works in the industry. I can tell you that the experience of demoing a microscope system to a potential customer often goes something like this: The potential customer prepares a typical biological specimen that they are familiar with from experience and then images of this specimen are captured with the demo system. These images are then compared to a similar set of images that were produced on another competing instrument (maybe - sometimes comparison images are not available). Usually there are statements made to the effect of "yes/no, the images on this instrument look better/worse, therefore I should buy/decline instrument A/B". I think anyone with a good imagination can figure out that this is a very subjective way to compare instrument performance and that there are a lot of potential pitfalls involved that might bias the assessment one way or another. I'm not saying this isn't a good method to judge an microscope's general imaging performance. Indeed, you should always take a prospective microscope system out for a "test drive", but a more quantitative test that doesn't solely depend on the opinion of how "good" an image looks would be better for everyone in my opinion. And Kurt, supposing a good set of text specimens and protocols were developed - how could we all come to agree on them? Who would set the standards and govern them for microscopy? Surely the industry players would have much to say (or dispute!) about that. Again, as stated above, there is a paramount need in microscopy for standardized test samples and procedures. It's a cause I certainly would be willing to devote time to were there an organized and concerted effort to make it happen. Curious to hear other people's thoughts on this. John Oreopoulos Staff Scientist Spectral Applied Research Inc. A Division of Andor Technology Richmond Hill, Ontario Canada www.spectral.ca On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** hi all, anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? is anyone aware of this issue ever having been discussed by NEMA? cheers kurt . Prof. Kurt I. Anderson Tumor Cell Migration Lab and Beatson Advanced Imaging Resource (BAIR) The Beatson Institute for Cancer Research Garscube Estate, Switchback Road, Glasgow G61 1BD • Direct Line +44 (0) 141 330 2864 • Fax +44 (0) 141 942 6521 [hidden email]<mailto:[hidden email]><mailto:[hidden email]> A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. ________________________________ This message is intended for the use of the person(s) to whom it may be addressed. It may contain information that is privileged, confidential, or otherwise protected from disclosure under applicable law. If you are not the intended recipient, any dissemination, distribution, copying, or use of this information is prohibited. If you have received this message in error, please permanently delete it and immediately notify the sender. Thank you. |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Well, that's very interesting. This is exactly the kind of test specimen I had in mind for fluorescence microscopy. I had not heard of this company before now. Thank you Glenn for pointing them out. I think I'll have to try using one of their test slides. What I find most interesting here is Argolight's ability to manufacture complex 3D structures with the fluorescent brush technology. The question then becomes - what's a good/challenging test specimen 3D pattern for confocal microscopy that everyone could all agree on? John Oreopoulos Staff Scientist Spectral Applied Research Inc. A Division of Andor Technology Richmond Hill, Ontario Canada www.spectral.ca On 2014-03-15, at 4:56 PM, Glenn Merrill-Skoloff wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > There’s a commercially available product that claims to meet most of the requirements John lists, below. Has anyone used this slide? > > http://argolight.com/product-micro/ > > My Best, > — Glenn > > Glenn Merrill-Skoloff > Division of Hemostasis and Thrombosis > Director, Intravital Microscopy Core > > 617-735-4040 (Office) > 617-735-4007 (Lab) > 617-735-4000 (Fax) > > > > From: John Oreopoulos <[hidden email]<mailto:[hidden email]>> > Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]>> > Date: Saturday, March 15, 2014 at 12:35 AM > To: "[hidden email]<mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]>> > Subject: Re: Confocal NEMA? > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Kurt, > > I'm not familiar with NEMA, but the general topic of testing/standardizing confocal sensitivity, resolution, and image quality is of great interest to me. If you search the confocal listserver archive, you'll find several postings about this. In theory, you could develop a standard test for confocal sensitivity, resolution, and image quality, but what should the test specimen be? Others have worked on this to some extent, and a few names that come to mind are Fred Brakenhoff, Claire Brown, Robert Zucker, and Mike Model. I recommend looking up their papers to see what sorts of test specimens have been proposed and tried in the past. Unfortunately, I don't think there is one single test specimen that can be used to determine all instrument parameters of interest (sensitivity, resolution - spatial, temporal, spectral-, and image quality, etc.). > > In my mind, an ideal test specimen for confocal fluorescence imaging would have the following properties: > > 1. Be cheap, readily available, and reproducible to a high degree > 2. Able to absorb and emit light over broad range of wavelengths > 3. Be portable and emit constant intensity (no photobleaching) perhaps even with a known number of photons under certain conditions > 4. Does not saturate easily, and has a linear response to excitation light power > 5. Has a known 3D structure of specific shapes/patterns/sizes with a high level of precision determined by some other imaging technique (electron microscopy for example). > 6. Has a refractive index close to water or glass > > Again, I don't think such a sample exists, but if you can think of one, let me know! I think one of the other problems here is: how would you define and quantify things like "image quality"? I'm curious how NEMA does this for PET. Spatial resolution is a bit more straightforward to quantify, although here too there are different ways to define resolution and ways to measure it. Sensitivity measurements are complicated by the need to keep many other imaging parameters constant when examining day-to-day performance/sensitivity of one instrument, or comparing the performance of two different instruments - see Jim Pawley's "39 steps". > > Your question also brings to mind a quote from another very well-written article on the topic which I never forget: > > "This complex relationship between qualitative and quantitative content of fluorescence images also expresses itself in the way that commercial instruments are assessed. On the one hand it is unavoidable that users initially are strongly influenced by the visual quality of the images presented by the instrument. On the other hand the longer term scientific value of the instrument depends crucially both on the quality of the visual information and the effectiveness with which it can be reduced for quantitative analysis... The increased complexity of a fluorescent image (in terms of the number of dimensions that can be measured), and the variability of sample preparation, makes it less easy to assess quantitatively the quality of the image. This may seem a surprising, or at least somewhat bleak, assessment. But consider this: In the 20 years or more since introduction of laser scanning confocal microscopes it has not been feasible, from published data, to assess how these systems compare (in terms of absolute units associated with measurements). No one can assert with confidence that instrument A in use in 1990 has better or worse sensitivity than instrument B operating in 2006. There is, therefore, a paramount need for standardized test samples and procedures for their use." > > -taken from: > Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization of fluorescence measurements from the instrument manufacturer's view standardization and quality assurance in fluorescence measurements ii. 2008. 6: p. 3-24. > > I find the second last statement above quite frustrating, especially from the point of view of someone who works in the industry. I can tell you that the experience of demoing a microscope system to a potential customer often goes something like this: The potential customer prepares a typical biological specimen that they are familiar with from experience and then images of this specimen are captured with the demo system. These images are then compared to a similar set of images that were produced on another competing instrument (maybe - sometimes comparison images are not available). Usually there are statements made to the effect of "yes/no, the images on this instrument look better/worse, therefore I should buy/decline instrument A/B". I think anyone with a good imagination can figure out that this is a very subjective way to compare instrument performance and that there are a lot of potential pitfalls involved that might bias the assessment one way or another. I'm not saying this isn't a good method to judge an microscope's general imaging performance. Indeed, you should always take a prospective microscope system out for a "test drive", but a more quantitative test that doesn't solely depend on the opinion of how "good" an image looks would be better for everyone in my opinion. > > And Kurt, supposing a good set of text specimens and protocols were developed - how could we all come to agree on them? Who would set the standards and govern them for microscopy? Surely the industry players would have much to say (or dispute!) about that. Again, as stated above, there is a paramount need in microscopy for standardized test samples and procedures. It's a cause I certainly would be willing to devote time to were there an organized and concerted effort to make it happen. > > Curious to hear other people's thoughts on this. > > > John Oreopoulos > Staff Scientist > Spectral Applied Research Inc. > A Division of Andor Technology > Richmond Hill, Ontario > Canada > www.spectral.ca > > > > On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > hi all, > anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. > they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. > see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. > recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. > among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. > my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? > is anyone aware of this issue ever having been discussed by NEMA? > cheers > kurt > . > Prof. Kurt I. Anderson > Tumor Cell Migration Lab and > Beatson Advanced Imaging Resource (BAIR) > The Beatson Institute for Cancer Research > Garscube Estate, Switchback Road, Glasgow G61 1BD > • Direct Line +44 (0) 141 330 2864 > • Fax +44 (0) 141 942 6521 > [hidden email]<mailto:[hidden email]><mailto:[hidden email]> > A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 > The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. > > > ________________________________ > This message is intended for the use of the person(s) to whom it may be addressed. It may contain information that is privileged, confidential, or otherwise protected from disclosure under applicable law. If you are not the intended recipient, any dissemination, distribution, copying, or use of this information is prohibited. If you have received this message in error, please permanently delete it and immediately notify the sender. Thank you. |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Here's a link to a recent article on the Argolight test slides: http://argolight.com/wp-content/uploads/2013/09/2013_Royon_Imaging-and-Microscopy.pdf I'd echo Glenn's question: Has anyone out there tried these slides yet? Very curious to know how well they meet the claims of the article. How do they prevent photobleaching I wonder, and for how long? John On 2014-03-15, at 11:43 PM, John Oreopoulos wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Well, that's very interesting. This is exactly the kind of test specimen I had in mind for fluorescence microscopy. I had not heard of this company before now. Thank you Glenn for pointing them out. I think I'll have to try using one of their test slides. What I find most interesting here is Argolight's ability to manufacture complex 3D structures with the fluorescent brush technology. The question then becomes - what's a good/challenging test specimen 3D pattern for confocal microscopy that everyone could all agree on? > > John Oreopoulos > Staff Scientist > Spectral Applied Research Inc. > A Division of Andor Technology > Richmond Hill, Ontario > Canada > www.spectral.ca > > > On 2014-03-15, at 4:56 PM, Glenn Merrill-Skoloff wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> There’s a commercially available product that claims to meet most of the requirements John lists, below. Has anyone used this slide? >> >> http://argolight.com/product-micro/ >> >> My Best, >> — Glenn >> >> Glenn Merrill-Skoloff >> Division of Hemostasis and Thrombosis >> Director, Intravital Microscopy Core >> >> 617-735-4040 (Office) >> 617-735-4007 (Lab) >> 617-735-4000 (Fax) >> >> >> >> From: John Oreopoulos <[hidden email]<mailto:[hidden email]>> >> Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]>> >> Date: Saturday, March 15, 2014 at 12:35 AM >> To: "[hidden email]<mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]>> >> Subject: Re: Confocal NEMA? >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Kurt, >> >> I'm not familiar with NEMA, but the general topic of testing/standardizing confocal sensitivity, resolution, and image quality is of great interest to me. If you search the confocal listserver archive, you'll find several postings about this. In theory, you could develop a standard test for confocal sensitivity, resolution, and image quality, but what should the test specimen be? Others have worked on this to some extent, and a few names that come to mind are Fred Brakenhoff, Claire Brown, Robert Zucker, and Mike Model. I recommend looking up their papers to see what sorts of test specimens have been proposed and tried in the past. Unfortunately, I don't think there is one single test specimen that can be used to determine all instrument parameters of interest (sensitivity, resolution - spatial, temporal, spectral-, and image quality, etc.). >> >> In my mind, an ideal test specimen for confocal fluorescence imaging would have the following properties: >> >> 1. Be cheap, readily available, and reproducible to a high degree >> 2. Able to absorb and emit light over broad range of wavelengths >> 3. Be portable and emit constant intensity (no photobleaching) perhaps even with a known number of photons under certain conditions >> 4. Does not saturate easily, and has a linear response to excitation light power >> 5. Has a known 3D structure of specific shapes/patterns/sizes with a high level of precision determined by some other imaging technique (electron microscopy for example). >> 6. Has a refractive index close to water or glass >> >> Again, I don't think such a sample exists, but if you can think of one, let me know! I think one of the other problems here is: how would you define and quantify things like "image quality"? I'm curious how NEMA does this for PET. Spatial resolution is a bit more straightforward to quantify, although here too there are different ways to define resolution and ways to measure it. Sensitivity measurements are complicated by the need to keep many other imaging parameters constant when examining day-to-day performance/sensitivity of one instrument, or comparing the performance of two different instruments - see Jim Pawley's "39 steps". >> >> Your question also brings to mind a quote from another very well-written article on the topic which I never forget: >> >> "This complex relationship between qualitative and quantitative content of fluorescence images also expresses itself in the way that commercial instruments are assessed. On the one hand it is unavoidable that users initially are strongly influenced by the visual quality of the images presented by the instrument. On the other hand the longer term scientific value of the instrument depends crucially both on the quality of the visual information and the effectiveness with which it can be reduced for quantitative analysis... The increased complexity of a fluorescent image (in terms of the number of dimensions that can be measured), and the variability of sample preparation, makes it less easy to assess quantitatively the quality of the image. This may seem a surprising, or at least somewhat bleak, assessment. But consider this: In the 20 years or more since introduction of laser scanning confocal microscopes it has not been feasible, from published data, to assess how these systems compare (in terms of absolute units associated with measurements). No one can assert with confidence that instrument A in use in 1990 has better or worse sensitivity than instrument B operating in 2006. There is, therefore, a paramount need for standardized test samples and procedures for their use." >> >> -taken from: >> Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization of fluorescence measurements from the instrument manufacturer's view standardization and quality assurance in fluorescence measurements ii. 2008. 6: p. 3-24. >> >> I find the second last statement above quite frustrating, especially from the point of view of someone who works in the industry. I can tell you that the experience of demoing a microscope system to a potential customer often goes something like this: The potential customer prepares a typical biological specimen that they are familiar with from experience and then images of this specimen are captured with the demo system. These images are then compared to a similar set of images that were produced on another competing instrument (maybe - sometimes comparison images are not available). Usually there are statements made to the effect of "yes/no, the images on this instrument look better/worse, therefore I should buy/decline instrument A/B". I think anyone with a good imagination can figure out that this is a very subjective way to compare instrument performance and that there are a lot of potential pitfalls involved that might bias the assessment one way or another. I'm not saying this isn't a good method to judge an microscope's general imaging performance. Indeed, you should always take a prospective microscope system out for a "test drive", but a more quantitative test that doesn't solely depend on the opinion of how "good" an image looks would be better for everyone in my opinion. >> >> And Kurt, supposing a good set of text specimens and protocols were developed - how could we all come to agree on them? Who would set the standards and govern them for microscopy? Surely the industry players would have much to say (or dispute!) about that. Again, as stated above, there is a paramount need in microscopy for standardized test samples and procedures. It's a cause I certainly would be willing to devote time to were there an organized and concerted effort to make it happen. >> >> Curious to hear other people's thoughts on this. >> >> >> John Oreopoulos >> Staff Scientist >> Spectral Applied Research Inc. >> A Division of Andor Technology >> Richmond Hill, Ontario >> Canada >> www.spectral.ca >> >> >> >> On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> hi all, >> anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. >> they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. >> see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. >> recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. >> among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. >> my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? >> is anyone aware of this issue ever having been discussed by NEMA? >> cheers >> kurt >> . >> Prof. Kurt I. Anderson >> Tumor Cell Migration Lab and >> Beatson Advanced Imaging Resource (BAIR) >> The Beatson Institute for Cancer Research >> Garscube Estate, Switchback Road, Glasgow G61 1BD >> • Direct Line +44 (0) 141 330 2864 >> • Fax +44 (0) 141 942 6521 >> [hidden email]<mailto:[hidden email]><mailto:[hidden email]> >> A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 >> The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. >> >> >> ________________________________ >> This message is intended for the use of the person(s) to whom it may be addressed. It may contain information that is privileged, confidential, or otherwise protected from disclosure under applicable law. If you are not the intended recipient, any dissemination, distribution, copying, or use of this information is prohibited. If you have received this message in error, please permanently delete it and immediately notify the sender. Thank you. |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** How much do these slides cost? Sent from my iPad > On Mar 15, 2014, at 11:28 PM, "John Oreopoulos" <[hidden email]> wrote: > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Here's a link to a recent article on the Argolight test slides: > > http://argolight.com/wp-content/uploads/2013/09/2013_Royon_Imaging-and-Microscopy.pdf > > I'd echo Glenn's question: Has anyone out there tried these slides yet? Very curious to know how well they meet the claims of the article. How do they prevent photobleaching I wonder, and for how long? > > John > > > >> On 2014-03-15, at 11:43 PM, John Oreopoulos wrote: >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Well, that's very interesting. This is exactly the kind of test specimen I had in mind for fluorescence microscopy. I had not heard of this company before now. Thank you Glenn for pointing them out. I think I'll have to try using one of their test slides. What I find most interesting here is Argolight's ability to manufacture complex 3D structures with the fluorescent brush technology. The question then becomes - what's a good/challenging test specimen 3D pattern for confocal microscopy that everyone could all agree on? >> >> John Oreopoulos >> Staff Scientist >> Spectral Applied Research Inc. >> A Division of Andor Technology >> Richmond Hill, Ontario >> Canada >> www.spectral.ca >> >> >>> On 2014-03-15, at 4:56 PM, Glenn Merrill-Skoloff wrote: >>> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> There’s a commercially available product that claims to meet most of the requirements John lists, below. Has anyone used this slide? >>> >>> http://argolight.com/product-micro/ >>> >>> My Best, >>> — Glenn >>> >>> Glenn Merrill-Skoloff >>> Division of Hemostasis and Thrombosis >>> Director, Intravital Microscopy Core >>> >>> 617-735-4040 (Office) >>> 617-735-4007 (Lab) >>> 617-735-4000 (Fax) >>> >>> >>> >>> From: John Oreopoulos <[hidden email]<mailto:[hidden email]>> >>> Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]>> >>> Date: Saturday, March 15, 2014 at 12:35 AM >>> To: "[hidden email]<mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]>> >>> Subject: Re: Confocal NEMA? >>> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> Kurt, >>> >>> I'm not familiar with NEMA, but the general topic of testing/standardizing confocal sensitivity, resolution, and image quality is of great interest to me. If you search the confocal listserver archive, you'll find several postings about this. In theory, you could develop a standard test for confocal sensitivity, resolution, and image quality, but what should the test specimen be? Others have worked on this to some extent, and a few names that come to mind are Fred Brakenhoff, Claire Brown, Robert Zucker, and Mike Model. I recommend looking up their papers to see what sorts of test specimens have been proposed and tried in the past. Unfortunately, I don't think there is one single test specimen that can be used to determine all instrument parameters of interest (sensitivity, resolution - spatial, temporal, spectral-, and image quality, etc.). >>> >>> In my mind, an ideal test specimen for confocal fluorescence imaging would have the following properties: >>> >>> 1. Be cheap, readily available, and reproducible to a high degree >>> 2. Able to absorb and emit light over broad range of wavelengths >>> 3. Be portable and emit constant intensity (no photobleaching) perhaps even with a known number of photons under certain conditions >>> 4. Does not saturate easily, and has a linear response to excitation light power >>> 5. Has a known 3D structure of specific shapes/patterns/sizes with a high level of precision determined by some other imaging technique (electron microscopy for example). >>> 6. Has a refractive index close to water or glass >>> >>> Again, I don't think such a sample exists, but if you can think of one, let me know! I think one of the other problems here is: how would you define and quantify things like "image quality"? I'm curious how NEMA does this for PET. Spatial resolution is a bit more straightforward to quantify, although here too there are different ways to define resolution and ways to measure it. Sensitivity measurements are complicated by the need to keep many other imaging parameters constant when examining day-to-day performance/sensitivity of one instrument, or comparing the performance of two different instruments - see Jim Pawley's "39 steps". >>> >>> Your question also brings to mind a quote from another very well-written article on the topic which I never forget: >>> >>> "This complex relationship between qualitative and quantitative content of fluorescence images also expresses itself in the way that commercial instruments are assessed. On the one hand it is unavoidable that users initially are strongly influenced by the visual quality of the images presented by the instrument. On the other hand the longer term scientific value of the instrument depends crucially both on the quality of the visual information and the effectiveness with which it can be reduced for quantitative analysis... The increased complexity of a fluorescent image (in terms of the number of dimensions that can be measured), and the variability of sample preparation, makes it less easy to assess quantitatively the quality of the image. This may seem a surprising, or at least somewhat bleak, assessment. But consider this: In the 20 years or more since introduction of laser scanning confocal microscopes it has not been feasible, from published data, to assess how these systems compare (in terms of absolute units associated with measurements). No one can assert with confidence that instrument A in use in 1990 has better or worse sensitivity than instrument B operating in 2006. There is, therefore, a paramount need for standardized test samples and procedures for their use." >>> >>> -taken from: >>> Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization of fluorescence measurements from the instrument manufacturer's view standardization and quality assurance in fluorescence measurements ii. 2008. 6: p. 3-24. >>> >>> I find the second last statement above quite frustrating, especially from the point of view of someone who works in the industry. I can tell you that the experience of demoing a microscope system to a potential customer often goes something like this: The potential customer prepares a typical biological specimen that they are familiar with from experience and then images of this specimen are captured with the demo system. These images are then compared to a similar set of images that were produced on another competing instrument (maybe - sometimes comparison images are not available). Usually there are statements made to the effect of "yes/no, the images on this instrument look better/worse, therefore I should buy/decline instrument A/B". I think anyone with a good imagination can figure out that this is a very subjective way to compare instrument performance and that there are a lot of potential pitfalls involved that might bias the assessment one way or another. I'm not saying this isn't a good method to judge an microscope's general imaging performance. Indeed, you should always take a prospective microscope system out for a "test drive", but a more quantitative test that doesn't solely depend on the opinion of how "good" an image looks would be better for everyone in my opinion. >>> >>> And Kurt, supposing a good set of text specimens and protocols were developed - how could we all come to agree on them? Who would set the standards and govern them for microscopy? Surely the industry players would have much to say (or dispute!) about that. Again, as stated above, there is a paramount need in microscopy for standardized test samples and procedures. It's a cause I certainly would be willing to devote time to were there an organized and concerted effort to make it happen. >>> >>> Curious to hear other people's thoughts on this. >>> >>> >>> John Oreopoulos >>> Staff Scientist >>> Spectral Applied Research Inc. >>> A Division of Andor Technology >>> Richmond Hill, Ontario >>> Canada >>> www.spectral.ca >>> >>> >>> >>> On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: >>> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> hi all, >>> anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. >>> they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. >>> see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. >>> recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. >>> among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. >>> my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? >>> is anyone aware of this issue ever having been discussed by NEMA? >>> cheers >>> kurt >>> . >>> Prof. Kurt I. Anderson >>> Tumor Cell Migration Lab and >>> Beatson Advanced Imaging Resource (BAIR) >>> The Beatson Institute for Cancer Research >>> Garscube Estate, Switchback Road, Glasgow G61 1BD >>> • Direct Line +44 (0) 141 330 2864 >>> • Fax +44 (0) 141 942 6521 >>> [hidden email]<mailto:[hidden email]><mailto:[hidden email]> >>> A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 >>> The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. >>> >>> >>> ________________________________ >>> This message is intended for the use of the person(s) to whom it may be addressed. It may contain information that is privileged, confidential, or otherwise protected from disclosure under applicable law. If you are not the intended recipient, any dissemination, distribution, copying, or use of this information is prohibited. If you have received this message in error, please permanently delete it and immediately notify the sender. Thank you. |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I had the standard slide quoted to me for 5000 euro last May. — Glenn Glenn Merrill-Skoloff Beth Israel Deaconess Medical Center Division of Hemostasis and Thrombosis Director, Intravital Microscopy Core 617-735-4040 (Office) 617-735-4007 (Lab) 617-735-4000 (Fax) From: <Phillips>, "Thomas E." <[hidden email]<mailto:[hidden email]>> Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]>> Date: Sunday, March 16, 2014 at 8:49 AM To: "[hidden email]<mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]>> Subject: Re: Confocal NEMA? ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** How much do these slides cost? Sent from my iPad On Mar 15, 2014, at 11:28 PM, "John Oreopoulos" <[hidden email]<mailto:[hidden email]>> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Here's a link to a recent article on the Argolight test slides: http://argolight.com/wp-content/uploads/2013/09/2013_Royon_Imaging-and-Microscopy.pdf I'd echo Glenn's question: Has anyone out there tried these slides yet? Very curious to know how well they meet the claims of the article. How do they prevent photobleaching I wonder, and for how long? John On 2014-03-15, at 11:43 PM, John Oreopoulos wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Well, that's very interesting. This is exactly the kind of test specimen I had in mind for fluorescence microscopy. I had not heard of this company before now. Thank you Glenn for pointing them out. I think I'll have to try using one of their test slides. What I find most interesting here is Argolight's ability to manufacture complex 3D structures with the fluorescent brush technology. The question then becomes - what's a good/challenging test specimen 3D pattern for confocal microscopy that everyone could all agree on? John Oreopoulos Staff Scientist Spectral Applied Research Inc. A Division of Andor Technology Richmond Hill, Ontario Canada www.spectral.ca On 2014-03-15, at 4:56 PM, Glenn Merrill-Skoloff wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** There’s a commercially available product that claims to meet most of the requirements John lists, below. Has anyone used this slide? http://argolight.com/product-micro/ My Best, — Glenn Glenn Merrill-Skoloff Division of Hemostasis and Thrombosis Director, Intravital Microscopy Core 617-735-4040 (Office) 617-735-4007 (Lab) 617-735-4000 (Fax) From: John Oreopoulos <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> Date: Saturday, March 15, 2014 at 12:35 AM To: "[hidden email]<mailto:[hidden email]><mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> Subject: Re: Confocal NEMA? ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Kurt, I'm not familiar with NEMA, but the general topic of testing/standardizing confocal sensitivity, resolution, and image quality is of great interest to me. If you search the confocal listserver archive, you'll find several postings about this. In theory, you could develop a standard test for confocal sensitivity, resolution, and image quality, but what should the test specimen be? Others have worked on this to some extent, and a few names that come to mind are Fred Brakenhoff, Claire Brown, Robert Zucker, and Mike Model. I recommend looking up their papers to see what sorts of test specimens have been proposed and tried in the past. Unfortunately, I don't think there is one single test specimen that can be used to determine all instrument parameters of interest (sensitivity, resolution - spatial, temporal, spectral-, and image quality, etc.). In my mind, an ideal test specimen for confocal fluorescence imaging would have the following properties: 1. Be cheap, readily available, and reproducible to a high degree 2. Able to absorb and emit light over broad range of wavelengths 3. Be portable and emit constant intensity (no photobleaching) perhaps even with a known number of photons under certain conditions 4. Does not saturate easily, and has a linear response to excitation light power 5. Has a known 3D structure of specific shapes/patterns/sizes with a high level of precision determined by some other imaging technique (electron microscopy for example). 6. Has a refractive index close to water or glass Again, I don't think such a sample exists, but if you can think of one, let me know! I think one of the other problems here is: how would you define and quantify things like "image quality"? I'm curious how NEMA does this for PET. Spatial resolution is a bit more straightforward to quantify, although here too there are different ways to define resolution and ways to measure it. Sensitivity measurements are complicated by the need to keep many other imaging parameters constant when examining day-to-day performance/sensitivity of one instrument, or comparing the performance of two different instruments - see Jim Pawley's "39 steps". Your question also brings to mind a quote from another very well-written article on the topic which I never forget: "This complex relationship between qualitative and quantitative content of fluorescence images also expresses itself in the way that commercial instruments are assessed. On the one hand it is unavoidable that users initially are strongly influenced by the visual quality of the images presented by the instrument. On the other hand the longer term scientific value of the instrument depends crucially both on the quality of the visual information and the effectiveness with which it can be reduced for quantitative analysis... The increased complexity of a fluorescent image (in terms of the number of dimensions that can be measured), and the variability of sample preparation, makes it less easy to assess quantitatively the quality of the image. This may seem a surprising, or at least somewhat bleak, assessment. But consider this: In the 20 years or more since introduction of laser scanning confocal microscopes it has not been feasible, from published data, to assess how these systems compare (in terms of absolute units associated with measurements). No one can assert with confidence that instrument A in use in 1990 has better or worse sensitivity than instrument B operating in 2006. There is, therefore, a paramount need for standardized test samples and procedures for their use." -taken from: Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization of fluorescence measurements from the instrument manufacturer's view standardization and quality assurance in fluorescence measurements ii. 2008. 6: p. 3-24. I find the second last statement above quite frustrating, especially from the point of view of someone who works in the industry. I can tell you that the experience of demoing a microscope system to a potential customer often goes something like this: The potential customer prepares a typical biological specimen that they are familiar with from experience and then images of this specimen are captured with the demo system. These images are then compared to a similar set of images that were produced on another competing instrument (maybe - sometimes comparison images are not available). Usually there are statements made to the effect of "yes/no, the images on this instrument look better/worse, therefore I should buy/decline instrument A/B". I think anyone with a good imagination can figure out that this is a very subjective way to compare instrument performance and that there are a lot of potential pitfalls involved that might bias the assessment one way or another. I'm not saying this isn't a good method to judge an microscope's general imaging performance. Indeed, you should always take a prospective microscope system out for a "test drive", but a more quantitative test that doesn't solely depend on the opinion of how "good" an image looks would be better for everyone in my opinion. And Kurt, supposing a good set of text specimens and protocols were developed - how could we all come to agree on them? Who would set the standards and govern them for microscopy? Surely the industry players would have much to say (or dispute!) about that. Again, as stated above, there is a paramount need in microscopy for standardized test samples and procedures. It's a cause I certainly would be willing to devote time to were there an organized and concerted effort to make it happen. Curious to hear other people's thoughts on this. John Oreopoulos Staff Scientist Spectral Applied Research Inc. A Division of Andor Technology Richmond Hill, Ontario Canada www.spectral.ca On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** hi all, anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? is anyone aware of this issue ever having been discussed by NEMA? cheers kurt . Prof. Kurt I. Anderson Tumor Cell Migration Lab and Beatson Advanced Imaging Resource (BAIR) The Beatson Institute for Cancer Research Garscube Estate, Switchback Road, Glasgow G61 1BD • Direct Line +44 (0) 141 330 2864 • Fax +44 (0) 141 942 6521 [hidden email]<mailto:[hidden email]><mailto:[hidden email]><mailto:[hidden email]> A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. ________________________________ This message is intended for the use of the person(s) to whom it may be addressed. It may contain information that is privileged, confidential, or otherwise protected from disclosure under applicable law. If you are not the intended recipient, any dissemination, distribution, copying, or use of this information is prohibited. If you have received this message in error, please permanently delete it and immediately notify the sender. Thank you. |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Ouch. Doesn't really satisfy the "cheap, and readily available" criterion, does it. John Oreopoulos Staff Scientist Spectral Applied Research Inc. A Division of Andor Technology Richmond Hill, Ontario Canada www.spectral.ca On 2014-03-16, at 2:31 PM, Glenn Merrill-Skoloff wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > I had the standard slide quoted to me for 5000 euro last May. > > — Glenn > > Glenn Merrill-Skoloff > Beth Israel Deaconess Medical Center > Division of Hemostasis and Thrombosis > Director, Intravital Microscopy Core > > > 617-735-4040 (Office) > 617-735-4007 (Lab) > 617-735-4000 (Fax) > > > > From: <Phillips>, "Thomas E." <[hidden email]<mailto:[hidden email]>> > Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]>> > Date: Sunday, March 16, 2014 at 8:49 AM > To: "[hidden email]<mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]>> > Subject: Re: Confocal NEMA? > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > How much do these slides cost? > > Sent from my iPad > > On Mar 15, 2014, at 11:28 PM, "John Oreopoulos" <[hidden email]<mailto:[hidden email]>> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > Here's a link to a recent article on the Argolight test slides: > http://argolight.com/wp-content/uploads/2013/09/2013_Royon_Imaging-and-Microscopy.pdf > I'd echo Glenn's question: Has anyone out there tried these slides yet? Very curious to know how well they meet the claims of the article. How do they prevent photobleaching I wonder, and for how long? > John > On 2014-03-15, at 11:43 PM, John Oreopoulos wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > Well, that's very interesting. This is exactly the kind of test specimen I had in mind for fluorescence microscopy. I had not heard of this company before now. Thank you Glenn for pointing them out. I think I'll have to try using one of their test slides. What I find most interesting here is Argolight's ability to manufacture complex 3D structures with the fluorescent brush technology. The question then becomes - what's a good/challenging test specimen 3D pattern for confocal microscopy that everyone could all agree on? > John Oreopoulos > Staff Scientist > Spectral Applied Research Inc. > A Division of Andor Technology > Richmond Hill, Ontario > Canada > www.spectral.ca > On 2014-03-15, at 4:56 PM, Glenn Merrill-Skoloff wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > There’s a commercially available product that claims to meet most of the requirements John lists, below. Has anyone used this slide? > http://argolight.com/product-micro/ > My Best, > — Glenn > Glenn Merrill-Skoloff > Division of Hemostasis and Thrombosis > Director, Intravital Microscopy Core > 617-735-4040 (Office) > 617-735-4007 (Lab) > 617-735-4000 (Fax) > From: John Oreopoulos <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> > Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> > Date: Saturday, March 15, 2014 at 12:35 AM > To: "[hidden email]<mailto:[hidden email]><mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> > Subject: Re: Confocal NEMA? > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > Kurt, > I'm not familiar with NEMA, but the general topic of testing/standardizing confocal sensitivity, resolution, and image quality is of great interest to me. If you search the confocal listserver archive, you'll find several postings about this. In theory, you could develop a standard test for confocal sensitivity, resolution, and image quality, but what should the test specimen be? Others have worked on this to some extent, and a few names that come to mind are Fred Brakenhoff, Claire Brown, Robert Zucker, and Mike Model. I recommend looking up their papers to see what sorts of test specimens have been proposed and tried in the past. Unfortunately, I don't think there is one single test specimen that can be used to determine all instrument parameters of interest (sensitivity, resolution - spatial, temporal, spectral-, and image quality, etc.). > In my mind, an ideal test specimen for confocal fluorescence imaging would have the following properties: > 1. Be cheap, readily available, and reproducible to a high degree > 2. Able to absorb and emit light over broad range of wavelengths > 3. Be portable and emit constant intensity (no photobleaching) perhaps even with a known number of photons under certain conditions > 4. Does not saturate easily, and has a linear response to excitation light power > 5. Has a known 3D structure of specific shapes/patterns/sizes with a high level of precision determined by some other imaging technique (electron microscopy for example). > 6. Has a refractive index close to water or glass > Again, I don't think such a sample exists, but if you can think of one, let me know! I think one of the other problems here is: how would you define and quantify things like "image quality"? I'm curious how NEMA does this for PET. Spatial resolution is a bit more straightforward to quantify, although here too there are different ways to define resolution and ways to measure it. Sensitivity measurements are complicated by the need to keep many other imaging parameters constant when examining day-to-day performance/sensitivity of one instrument, or comparing the performance of two different instruments - see Jim Pawley's "39 steps". > Your question also brings to mind a quote from another very well-written article on the topic which I never forget: > "This complex relationship between qualitative and quantitative content of fluorescence images also expresses itself in the way that commercial instruments are assessed. On the one hand it is unavoidable that users initially are strongly influenced by the visual quality of the images presented by the instrument. On the other hand the longer term scientific value of the instrument depends crucially both on the quality of the visual information and the effectiveness with which it can be reduced for quantitative analysis... The increased complexity of a fluorescent image (in terms of the number of dimensions that can be measured), and the variability of sample preparation, makes it less easy to assess quantitatively the quality of the image. This may seem a surprising, or at least somewhat bleak, assessment. But consider this: In the 20 years or more since introduction of laser scanning confocal microscopes it has not been feasible, from published data, to assess how these systems compare (in terms of absolute units associated with measurements). No one can assert with confidence that instrument A in use in 1990 has better or worse sensitivity than instrument B operating in 2006. There is, therefore, a paramount need for standardized test samples and procedures for their use." > -taken from: > Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization of fluorescence measurements from the instrument manufacturer's view standardization and quality assurance in fluorescence measurements ii. 2008. 6: p. 3-24. > I find the second last statement above quite frustrating, especially from the point of view of someone who works in the industry. I can tell you that the experience of demoing a microscope system to a potential customer often goes something like this: The potential customer prepares a typical biological specimen that they are familiar with from experience and then images of this specimen are captured with the demo system. These images are then compared to a similar set of images that were produced on another competing instrument (maybe - sometimes comparison images are not available). Usually there are statements made to the effect of "yes/no, the images on this instrument look better/worse, therefore I should buy/decline instrument A/B". I think anyone with a good imagination can figure out that this is a very subjective way to compare instrument performance and that there are a lot of potential pitfalls involved that might bias the assessment one way or another. I'm not saying this isn't a good method to judge an microscope's general imaging performance. Indeed, you should always take a prospective microscope system out for a "test drive", but a more quantitative test that doesn't solely depend on the opinion of how "good" an image looks would be better for everyone in my opinion. > And Kurt, supposing a good set of text specimens and protocols were developed - how could we all come to agree on them? Who would set the standards and govern them for microscopy? Surely the industry players would have much to say (or dispute!) about that. Again, as stated above, there is a paramount need in microscopy for standardized test samples and procedures. It's a cause I certainly would be willing to devote time to were there an organized and concerted effort to make it happen. > Curious to hear other people's thoughts on this. > John Oreopoulos > Staff Scientist > Spectral Applied Research Inc. > A Division of Andor Technology > Richmond Hill, Ontario > Canada > www.spectral.ca > On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > hi all, > anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. > they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. > see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. > recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. > among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. > my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? > is anyone aware of this issue ever having been discussed by NEMA? > cheers > kurt > . > Prof. Kurt I. Anderson > Tumor Cell Migration Lab and > Beatson Advanced Imaging Resource (BAIR) > The Beatson Institute for Cancer Research > Garscube Estate, Switchback Road, Glasgow G61 1BD > • Direct Line +44 (0) 141 330 2864 > • Fax +44 (0) 141 942 6521 > [hidden email]<mailto:[hidden email]><mailto:[hidden email]><mailto:[hidden email]> > A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 > The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. > ________________________________ > This message is intended for the use of the person(s) to whom it may be addressed. It may contain information that is privileged, confidential, or otherwise protected from disclosure under applicable law. If you are not the intended recipient, any dissemination, distribution, copying, or use of this information is prohibited. If you have received this message in error, please permanently delete it and immediately notify the sender. Thank you. |
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In reply to this post by Kurt Anderson-3
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** thanks glen and john for your thoughtful replies, i agree that there is no such thing as a perfect test specimen. the point, i suppose, is to agree on a set of specimens which are “useable” and on methods for evaluating them. in that way you can achieve a benchmark, or standard, which can be used to objectively compare the performance of various instruments. there are various scientific organisations, which you could imagine undertaking to establish confocal microscopy standards (RMS, ELMI, EuroBioImaging...). but its interesting that NEMA seem to have done this across a wide range of equipment categories united only by having something to do with electricity. it appears that no one is aware of the idea for confocal microscope performance standards ever having been raised with NEMA, so my next query will be to them to find out what goes into setting up a standard such as NU4-2008. cheers kurt . ps: personally, one of my favourite test samples is a drop of aqueous FITC beneath a coverslip. doesn’t bleach appreciably and stepping through from the coverslip into the dye is a great way to estimate optical section thickness. but i think the point really is that On 14 Mar 2014, at 17:14, Kurt Anderson <[hidden email]<mailto:[hidden email]>> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** hi all, anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? is anyone aware of this issue ever having been discussed by NEMA? cheers kurt . Prof. Kurt I. Anderson Tumor Cell Migration Lab and Beatson Advanced Imaging Resource (BAIR) The Beatson Institute for Cancer Research Garscube Estate, Switchback Road, Glasgow G61 1BD • Direct Line +44 (0) 141 330 2864 • Fax +44 (0) 141 942 6521 [hidden email]<mailto:[hidden email]><mailto:[hidden email]> A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. Prof. Kurt I. Anderson Tumor Cell Migration Lab and Beatson Advanced Imaging Resource (BAIR) The Beatson Institute for Cancer Research Garscube Estate, Switchback Road, Glasgow G61 1BD • Direct Line +44 (0) 141 330 2864 • Fax +44 (0) 141 942 6521 [hidden email]<mailto:[hidden email]> A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. |
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In reply to this post by Kurt Anderson-3
If one of these calibration slides was made available for a so called road show would the community be willing to collect measurements and then post them to a public web page or forum? The slide could then move around the world from facility to facility with the goal of collecting demographics from different techniques and instruments.
Robin Battye, M.Sc., Ph. D. Product Manager/ Technical Sales Specialist Mobile: (416) 302-5934 Spectral Applied Research A Division of Andor Technoloy 9078 Leslie St., Unit 11 Richmond Hill, Ontario L4B 3L8 Office: (905) 326-5040 ext. 406 Fax: (905) 326-5041 www.spectral.ca We believe that performance comes before Compromise. -------- Original message -------- From: John Oreopoulos <[hidden email]> Date: 16-03-2014 15:27 (GMT-05:00) To: [hidden email] Subject: Re: Confocal NEMA? ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Ouch. Doesn't really satisfy the "cheap, and readily available" criterion, does it. John Oreopoulos Staff Scientist Spectral Applied Research Inc. A Division of Andor Technology Richmond Hill, Ontario Canada www.spectral.ca On 2014-03-16, at 2:31 PM, Glenn Merrill-Skoloff wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > I had the standard slide quoted to me for 5000 euro last May. > > — Glenn > > Glenn Merrill-Skoloff > Beth Israel Deaconess Medical Center > Division of Hemostasis and Thrombosis > Director, Intravital Microscopy Core > > > 617-735-4040 (Office) > 617-735-4007 (Lab) > 617-735-4000 (Fax) > > > > From: <Phillips>, "Thomas E." <[hidden email]<mailto:[hidden email]>> > Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]>> > Date: Sunday, March 16, 2014 at 8:49 AM > To: "[hidden email]<mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]>> > Subject: Re: Confocal NEMA? > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > How much do these slides cost? > > Sent from my iPad > > On Mar 15, 2014, at 11:28 PM, "John Oreopoulos" <[hidden email]<mailto:[hidden email]>> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > Here's a link to a recent article on the Argolight test slides: > http://argolight.com/wp-content/uploads/2013/09/2013_Royon_Imaging-and-Microscopy.pdf > I'd echo Glenn's question: Has anyone out there tried these slides yet? Very curious to know how well they meet the claims of the article. How do they prevent photobleaching I wonder, and for how long? > John > On 2014-03-15, at 11:43 PM, John Oreopoulos wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > Well, that's very interesting. This is exactly the kind of test specimen I had in mind for fluorescence microscopy. I had not heard of this company before now. Thank you Glenn for pointing them out. I think I'll have to try using one of their test slides. What I find most interesting here is Argolight's ability to manufacture complex 3D structures with the fluorescent brush technology. The question then becomes - what's a good/challenging test specimen 3D pattern for confocal microscopy that everyone could all agree on? > John Oreopoulos > Staff Scientist > Spectral Applied Research Inc. > A Division of Andor Technology > Richmond Hill, Ontario > Canada > www.spectral.ca > On 2014-03-15, at 4:56 PM, Glenn Merrill-Skoloff wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > There’s a commercially available product that claims to meet most of the requirements John lists, below. Has anyone used this slide? > http://argolight.com/product-micro/ > My Best, > — Glenn > Glenn Merrill-Skoloff > Division of Hemostasis and Thrombosis > Director, Intravital Microscopy Core > 617-735-4040 (Office) > 617-735-4007 (Lab) > 617-735-4000 (Fax) > From: John Oreopoulos <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> > Reply-To: Confocal Microscopy List <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> > Date: Saturday, March 15, 2014 at 12:35 AM > To: "[hidden email]<mailto:[hidden email]><mailto:[hidden email]>" <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> > Subject: Re: Confocal NEMA? > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > Kurt, > I'm not familiar with NEMA, but the general topic of testing/standardizing confocal sensitivity, resolution, and image quality is of great interest to me. If you search the confocal listserver archive, you'll find several postings about this. In theory, you could develop a standard test for confocal sensitivity, resolution, and image quality, but what should the test specimen be? Others have worked on this to some extent, and a few names that come to mind are Fred Brakenhoff, Claire Brown, Robert Zucker, and Mike Model. I recommend looking up their papers to see what sorts of test specimens have been proposed and tried in the past. Unfortunately, I don't think there is one single test specimen that can be used to determine all instrument parameters of interest (sensitivity, resolution - spatial, temporal, spectral-, and image quality, etc.). > In my mind, an ideal test specimen for confocal fluorescence imaging would have the following properties: > 1. Be cheap, readily available, and reproducible to a high degree > 2. Able to absorb and emit light over broad range of wavelengths > 3. Be portable and emit constant intensity (no photobleaching) perhaps even with a known number of photons under certain conditions > 4. Does not saturate easily, and has a linear response to excitation light power > 5. Has a known 3D structure of specific shapes/patterns/sizes with a high level of precision determined by some other imaging technique (electron microscopy for example). > 6. Has a refractive index close to water or glass > Again, I don't think such a sample exists, but if you can think of one, let me know! I think one of the other problems here is: how would you define and quantify things like "image quality"? I'm curious how NEMA does this for PET. Spatial resolution is a bit more straightforward to quantify, although here too there are different ways to define resolution and ways to measure it. Sensitivity measurements are complicated by the need to keep many other imaging parameters constant when examining day-to-day performance/sensitivity of one instrument, or comparing the performance of two different instruments - see Jim Pawley's "39 steps". > Your question also brings to mind a quote from another very well-written article on the topic which I never forget: > "This complex relationship between qualitative and quantitative content of fluorescence images also expresses itself in the way that commercial instruments are assessed. On the one hand it is unavoidable that users initially are strongly influenced by the visual quality of the images presented by the instrument. On the other hand the longer term scientific value of the instrument depends crucially both on the quality of the visual information and the effectiveness with which it can be reduced for quantitative analysis... The increased complexity of a fluorescent image (in terms of the number of dimensions that can be measured), and the variability of sample preparation, makes it less easy to assess quantitatively the quality of the image. This may seem a surprising, or at least somewhat bleak, assessment. But consider this: In the 20 years or more since introduction of laser scanning confocal microscopes it has not been feasible, from published data, to assess how these systems compare (in terms of absolute units associated with measurements). No one can assert with confidence that instrument A in use in 1990 has better or worse sensitivity than instrument B operating in 2006. There is, therefore, a paramount need for standardized test samples and procedures for their use." > -taken from: > Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization of fluorescence measurements from the instrument manufacturer's view standardization and quality assurance in fluorescence measurements ii. 2008. 6: p. 3-24. > I find the second last statement above quite frustrating, especially from the point of view of someone who works in the industry. I can tell you that the experience of demoing a microscope system to a potential customer often goes something like this: The potential customer prepares a typical biological specimen that they are familiar with from experience and then images of this specimen are captured with the demo system. These images are then compared to a similar set of images that were produced on another competing instrument (maybe - sometimes comparison images are not available). Usually there are statements made to the effect of "yes/no, the images on this instrument look better/worse, therefore I should buy/decline instrument A/B". I think anyone with a good imagination can figure out that this is a very subjective way to compare instrument performance and that there are a lot of potential pitfalls involved that might bias the assessment one way or another. I'm not saying this isn't a good method to judge an microscope's general imaging performance. Indeed, you should always take a prospective microscope system out for a "test drive", but a more quantitative test that doesn't solely depend on the opinion of how "good" an image looks would be better for everyone in my opinion. > And Kurt, supposing a good set of text specimens and protocols were developed - how could we all come to agree on them? Who would set the standards and govern them for microscopy? Surely the industry players would have much to say (or dispute!) about that. Again, as stated above, there is a paramount need in microscopy for standardized test samples and procedures. It's a cause I certainly would be willing to devote time to were there an organized and concerted effort to make it happen. > Curious to hear other people's thoughts on this. > John Oreopoulos > Staff Scientist > Spectral Applied Research Inc. > A Division of Andor Technology > Richmond Hill, Ontario > Canada > www.spectral.ca > On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > hi all, > anyone who has worked in clinical or pre-clinical imaging will be familiar with the National Electrical Manufacturer’s Association, or NEMA. > they are like a north american DIN, in that they set standards for the specification and performance of electrical equipment ranging from conduit to clinical PET scanners. > see here<http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an interesting list of standards. > recently i’ve been involved in using NEMA protocol NU4-2008 to characterise a pre-clinical PET scanner. > among other things, the protocol specifies the samples and methods to be used for quantification of the instrument sensitivity, resolution, and image quality. > my naive question for a friday afternoon is this: why dont we have a NEMA standard for confocal microscopes? > is anyone aware of this issue ever having been discussed by NEMA? > cheers > kurt > . > Prof. Kurt I. Anderson > Tumor Cell Migration Lab and > Beatson Advanced Imaging Resource (BAIR) > The Beatson Institute for Cancer Research > Garscube Estate, Switchback Road, Glasgow G61 1BD > • Direct Line +44 (0) 141 330 2864 > • Fax +44 (0) 141 942 6521 > [hidden email]<mailto:[hidden email]><mailto:[hidden email]><mailto:[hidden email]> > A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106 > The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us. > ________________________________ > This message is intended for the use of the person(s) to whom it may be addressed. It may contain information that is privileged, confidential, or otherwise protected from disclosure under applicable law. If you are not the intended recipient, any dissemination, distribution, copying, or use of this information is prohibited. If you have received this message in error, please permanently delete it and immediately notify the sender. Thank you. |
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In reply to this post by John Oreopoulos
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** In November, I got quotes for 3000 and 4000 Euro, depending on the type of slide. I guess the price is probably justified by high production costs, but that won't help increasing their distribution. I also would be interested in reports on actual experience with these slides. Steffen Am 16.03.2014 20:27, schrieb John Oreopoulos: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Ouch. Doesn't really satisfy the "cheap, and readily available" criterion, does it. > > John Oreopoulos > Staff Scientist > Spectral Applied Research Inc. > A Division of Andor Technology > Richmond Hill, Ontario > Canada > www.spectral.ca > > > > On 2014-03-16, at 2:31 PM, Glenn Merrill-Skoloff wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> I had the standard slide quoted to me for 5000 euro last May. >> >> — Glenn >> >> Glenn Merrill-Skoloff >> Beth Israel Deaconess Medical Center >> Division of Hemostasis and Thrombosis >> Director, Intravital Microscopy Core >> >> >> 617-735-4040 (Office) >> 617-735-4007 (Lab) >> 617-735-4000 (Fax) -- ------------------------------------------------------------ Steffen Dietzel, PD Dr. rer. nat Ludwig-Maximilians-Universität München Walter-Brendel-Zentrum für experimentelle Medizin (WBex) Head of light microscopy Mail room: Marchioninistr. 15, D-81377 München Building location: Marchioninistr. 27, München-Großhadern |
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In reply to this post by John Oreopoulos
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** But isn't that exactly what the ABRF is trying to establish? http://www.ncbi.nlm.nih.gov/pubmed/21477410 http://www.abrf.org/index.cfm/group.show/LightMicroscopyResearchGroup.54.htm#membrs _____________________________________ Philippe Laissue, PhD, Bioimaging Manager School of Biological Sciences, Room 4.17 University of Essex, Colchester CO4 3SQ, UK (0044) 01206 872246 / (0044) 07842 676 456 [hidden email] privatewww.essex.ac.uk/~plaissue On 16 March 2014 03:43, John Oreopoulos <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Well, that's very interesting. This is exactly the kind of test specimen I > had in mind for fluorescence microscopy. I had not heard of this company > before now. Thank you Glenn for pointing them out. I think I'll have to try > using one of their test slides. What I find most interesting here is > Argolight's ability to manufacture complex 3D structures with the > fluorescent brush technology. The question then becomes - what's a > good/challenging test specimen 3D pattern for confocal microscopy that > everyone could all agree on? > > John Oreopoulos > Staff Scientist > Spectral Applied Research Inc. > A Division of Andor Technology > Richmond Hill, Ontario > Canada > www.spectral.ca > > > On 2014-03-15, at 4:56 PM, Glenn Merrill-Skoloff wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > There's a commercially available product that claims to meet most of the > requirements John lists, below. Has anyone used this slide? > > > > http://argolight.com/product-micro/ > > > > My Best, > > -- Glenn > > > > Glenn Merrill-Skoloff > > Division of Hemostasis and Thrombosis > > Director, Intravital Microscopy Core > > > > 617-735-4040 (Office) > > 617-735-4007 (Lab) > > 617-735-4000 (Fax) > > > > > > > > From: John Oreopoulos <[hidden email]<mailto: > [hidden email]>> > > Reply-To: Confocal Microscopy List <[hidden email] > <mailto:[hidden email]>> > > Date: Saturday, March 15, 2014 at 12:35 AM > > To: "[hidden email]<mailto: > [hidden email]>" <[hidden email] > <mailto:[hidden email]>> > > Subject: Re: Confocal NEMA? > > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Kurt, > > > > I'm not familiar with NEMA, but the general topic of > testing/standardizing confocal sensitivity, resolution, and image quality > is of great interest to me. If you search the confocal listserver archive, > you'll find several postings about this. In theory, you could develop a > standard test for confocal sensitivity, resolution, and image quality, but > what should the test specimen be? Others have worked on this to some > extent, and a few names that come to mind are Fred Brakenhoff, Claire > Brown, Robert Zucker, and Mike Model. I recommend looking up their papers > to see what sorts of test specimens have been proposed and tried in the > past. Unfortunately, I don't think there is one single test specimen that > can be used to determine all instrument parameters of interest > (sensitivity, resolution - spatial, temporal, spectral-, and image quality, > etc.). > > > > In my mind, an ideal test specimen for confocal fluorescence imaging > would have the following properties: > > > > 1. Be cheap, readily available, and reproducible to a high degree > > 2. Able to absorb and emit light over broad range of wavelengths > > 3. Be portable and emit constant intensity (no photobleaching) perhaps > even with a known number of photons under certain conditions > > 4. Does not saturate easily, and has a linear response to excitation > light power > > 5. Has a known 3D structure of specific shapes/patterns/sizes with a > high level of precision determined by some other imaging technique > (electron microscopy for example). > > 6. Has a refractive index close to water or glass > > > > Again, I don't think such a sample exists, but if you can think of one, > let me know! I think one of the other problems here is: how would you > define and quantify things like "image quality"? I'm curious how NEMA does > this for PET. Spatial resolution is a bit more straightforward to quantify, > although here too there are different ways to define resolution and ways to > measure it. Sensitivity measurements are complicated by the need to keep > many other imaging parameters constant when examining day-to-day > performance/sensitivity of one instrument, or comparing the performance of > two different instruments - see Jim Pawley's "39 steps". > > > > Your question also brings to mind a quote from another very well-written > article on the topic which I never forget: > > > > "This complex relationship between qualitative and quantitative content > of fluorescence images also expresses itself in the way that commercial > instruments are assessed. On the one hand it is unavoidable that users > initially are strongly influenced by the visual quality of the images > presented by the instrument. On the other hand the longer term scientific > value of the instrument depends crucially both on the quality of the visual > information and the effectiveness with which it can be reduced for > quantitative analysis... The increased complexity of a fluorescent image > (in terms of the number of dimensions that can be measured), and the > variability of sample preparation, makes it less easy to assess > quantitatively the quality of the image. This may seem a surprising, or at > least somewhat bleak, assessment. But consider this: In the 20 years or > more since introduction of laser scanning confocal microscopes it has not > been feasible, from published data, to assess how these systems compare (in > terms of absolute units associated with measurements). No one can assert > with confidence that instrument A in use in 1990 has better or worse > sensitivity than instrument B operating in 2006. There is, therefore, a > paramount need for standardized test samples and procedures for their use." > > > > -taken from: > > Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization > of fluorescence measurements from the instrument manufacturer's view > standardization and quality assurance in fluorescence measurements ii. > 2008. 6: p. 3-24. > > > > I find the second last statement above quite frustrating, especially > from the point of view of someone who works in the industry. I can tell you > that the experience of demoing a microscope system to a potential customer > often goes something like this: The potential customer prepares a typical > biological specimen that they are familiar with from experience and then > images of this specimen are captured with the demo system. These images are > then compared to a similar set of images that were produced on another > competing instrument (maybe - sometimes comparison images are not > available). Usually there are statements made to the effect of "yes/no, the > images on this instrument look better/worse, therefore I should buy/decline > instrument A/B". I think anyone with a good imagination can figure out that > this is a very subjective way to compare instrument performance and that > there are a lot of potential pitfalls involved that might bias the > assessment one way or another. I'm not saying this isn't a good method to > judge an microscope's general imaging performance. Indeed, you should > always take a prospective microscope system out for a "test drive", but a > more quantitative test that doesn't solely depend on the opinion of how > "good" an image looks would be better for everyone in my opinion. > > > > And Kurt, supposing a good set of text specimens and protocols were > developed - how could we all come to agree on them? Who would set the > standards and govern them for microscopy? Surely the industry players would > have much to say (or dispute!) about that. Again, as stated above, there is > a paramount need in microscopy for standardized test samples and > procedures. It's a cause I certainly would be willing to devote time to > were there an organized and concerted effort to make it happen. > > > > Curious to hear other people's thoughts on this. > > > > > > John Oreopoulos > > Staff Scientist > > Spectral Applied Research Inc. > > A Division of Andor Technology > > Richmond Hill, Ontario > > Canada > > www.spectral.ca > > > > > > > > On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: > > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > hi all, > > anyone who has worked in clinical or pre-clinical imaging will be > familiar with the National Electrical Manufacturer's Association, or NEMA. > > they are like a north american DIN, in that they set standards for the > specification and performance of electrical equipment ranging from conduit > to clinical PET scanners. > > see here< > http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an > interesting list of standards. > > recently i've been involved in using NEMA protocol NU4-2008 to > characterise a pre-clinical PET scanner. > > among other things, the protocol specifies the samples and methods to be > used for quantification of the instrument sensitivity, resolution, and > image quality. > > my naive question for a friday afternoon is this: why dont we have a > NEMA standard for confocal microscopes? > > is anyone aware of this issue ever having been discussed by NEMA? > > cheers > > kurt > > . > > Prof. Kurt I. Anderson > > Tumor Cell Migration Lab and > > Beatson Advanced Imaging Resource (BAIR) > > The Beatson Institute for Cancer Research > > Garscube Estate, Switchback Road, Glasgow G61 1BD > > * Direct Line +44 (0) 141 330 2864 > > * Fax +44 (0) 141 942 6521 > > [hidden email]<mailto:[hidden email] > ><mailto:[hidden email]> > > A company limited by guarantee; Registered in Scotland No 84170; A > Registered Scottish Charity No SC006106 > > The information contained in this communication may be privileged and > confidential and is intended for the exclusive use of the addressee > designated above. If you are not the addressee, any disclosure, > reproduction, distribution or other dissemination or use of this > communication is strictly prohibited. If you have received this > transmission in error please would you be kind enough to inform us. > > > > > > ________________________________ > > This message is intended for the use of the person(s) to whom it may be > addressed. It may contain information that is privileged, confidential, or > otherwise protected from disclosure under applicable law. If you are not > the intended recipient, any dissemination, distribution, copying, or use of > this information is prohibited. If you have received this message in error, > please permanently delete it and immediately notify the sender. Thank you. > |
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*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Yes, I would agree that the ABRF has made the greatest efforts in trying to measure microscope imaging performance on a somewhat large scale and develop imaging diagnostic protocols/standards. Their two recent publications are evidence of that: Stack, R. F., Bayles, C. J., Girard, A. M., Martin, K., Opansky, C., Schulz, K., et al. (2011). Quality assurance testing for modern optical imaging systems. Microscopy and Microanalysis, 17, 598-606. Cole, R. W., Thibault, M., Bayles, C. J., Eason, B., Girard, A. M., Jinadasa, T., et al. (2013). International test results for objective lens quality, resolution, spectral accuracy and spectral separation for confocal laser scanning microscopes. Microscopy and Microanalysis, 19, 1653-1668. They would be a logical organization to figure this sort of thing out. I suppose the RMS or a similar organization would also be up to the task and have the right resources for this. The acceptance of any kind of imaging standard (for example, see the USAF resolution target: http://en.wikipedia.org/wiki/1951_USAF_resolution_test_chart) would require some level of "metrological traceability" or validation as is described by NIST: http://www.nist.gov/traceability/nist_traceability_policy_external.cfm Again, it's not quite clear to me who the NEMA equivalent of the microscopy world is or if even such a body exists. What's interesting about NEMA is that its main members are big electrical companies like GE, 3M, and Siemans, so clearly there was an effort on the electrical industry's part to establish these electrical standards: http://www.nema.org/About/History/Pages/Model-of-Industrial-Self-Government.aspx I think ISO is the largest standardization organization I'm aware of: http://en.wikipedia.org/wiki/International_Organization_for_Standardization Are there any fluorescence microscopy or imaging ISO standards? John Oreopoulos Staff Scientist Spectral Applied Research Inc. A Division of Andor Technology Richmond Hill, Ontario Canada www.spectral.ca On 2014-03-17, at 7:00 AM, phil laissue wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > But isn't that exactly what the ABRF is trying to establish? > > http://www.ncbi.nlm.nih.gov/pubmed/21477410 > http://www.abrf.org/index.cfm/group.show/LightMicroscopyResearchGroup.54.htm#membrs > > > _____________________________________ > Philippe Laissue, PhD, Bioimaging Manager > School of Biological Sciences, Room 4.17 > University of Essex, Colchester CO4 3SQ, UK > (0044) 01206 872246 / (0044) 07842 676 456 > [hidden email] > privatewww.essex.ac.uk/~plaissue > > > On 16 March 2014 03:43, John Oreopoulos <[hidden email]> wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Well, that's very interesting. This is exactly the kind of test specimen I >> had in mind for fluorescence microscopy. I had not heard of this company >> before now. Thank you Glenn for pointing them out. I think I'll have to try >> using one of their test slides. What I find most interesting here is >> Argolight's ability to manufacture complex 3D structures with the >> fluorescent brush technology. The question then becomes - what's a >> good/challenging test specimen 3D pattern for confocal microscopy that >> everyone could all agree on? >> >> John Oreopoulos >> Staff Scientist >> Spectral Applied Research Inc. >> A Division of Andor Technology >> Richmond Hill, Ontario >> Canada >> www.spectral.ca >> >> >> On 2014-03-15, at 4:56 PM, Glenn Merrill-Skoloff wrote: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> There's a commercially available product that claims to meet most of the >> requirements John lists, below. Has anyone used this slide? >>> >>> http://argolight.com/product-micro/ >>> >>> My Best, >>> -- Glenn >>> >>> Glenn Merrill-Skoloff >>> Division of Hemostasis and Thrombosis >>> Director, Intravital Microscopy Core >>> >>> 617-735-4040 (Office) >>> 617-735-4007 (Lab) >>> 617-735-4000 (Fax) >>> >>> >>> >>> From: John Oreopoulos <[hidden email]<mailto: >> [hidden email]>> >>> Reply-To: Confocal Microscopy List <[hidden email] >> <mailto:[hidden email]>> >>> Date: Saturday, March 15, 2014 at 12:35 AM >>> To: "[hidden email]<mailto: >> [hidden email]>" <[hidden email] >> <mailto:[hidden email]>> >>> Subject: Re: Confocal NEMA? >>> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> Kurt, >>> >>> I'm not familiar with NEMA, but the general topic of >> testing/standardizing confocal sensitivity, resolution, and image quality >> is of great interest to me. If you search the confocal listserver archive, >> you'll find several postings about this. In theory, you could develop a >> standard test for confocal sensitivity, resolution, and image quality, but >> what should the test specimen be? Others have worked on this to some >> extent, and a few names that come to mind are Fred Brakenhoff, Claire >> Brown, Robert Zucker, and Mike Model. I recommend looking up their papers >> to see what sorts of test specimens have been proposed and tried in the >> past. Unfortunately, I don't think there is one single test specimen that >> can be used to determine all instrument parameters of interest >> (sensitivity, resolution - spatial, temporal, spectral-, and image quality, >> etc.). >>> >>> In my mind, an ideal test specimen for confocal fluorescence imaging >> would have the following properties: >>> >>> 1. Be cheap, readily available, and reproducible to a high degree >>> 2. Able to absorb and emit light over broad range of wavelengths >>> 3. Be portable and emit constant intensity (no photobleaching) perhaps >> even with a known number of photons under certain conditions >>> 4. Does not saturate easily, and has a linear response to excitation >> light power >>> 5. Has a known 3D structure of specific shapes/patterns/sizes with a >> high level of precision determined by some other imaging technique >> (electron microscopy for example). >>> 6. Has a refractive index close to water or glass >>> >>> Again, I don't think such a sample exists, but if you can think of one, >> let me know! I think one of the other problems here is: how would you >> define and quantify things like "image quality"? I'm curious how NEMA does >> this for PET. Spatial resolution is a bit more straightforward to quantify, >> although here too there are different ways to define resolution and ways to >> measure it. Sensitivity measurements are complicated by the need to keep >> many other imaging parameters constant when examining day-to-day >> performance/sensitivity of one instrument, or comparing the performance of >> two different instruments - see Jim Pawley's "39 steps". >>> >>> Your question also brings to mind a quote from another very well-written >> article on the topic which I never forget: >>> >>> "This complex relationship between qualitative and quantitative content >> of fluorescence images also expresses itself in the way that commercial >> instruments are assessed. On the one hand it is unavoidable that users >> initially are strongly influenced by the visual quality of the images >> presented by the instrument. On the other hand the longer term scientific >> value of the instrument depends crucially both on the quality of the visual >> information and the effectiveness with which it can be reduced for >> quantitative analysis... The increased complexity of a fluorescent image >> (in terms of the number of dimensions that can be measured), and the >> variability of sample preparation, makes it less easy to assess >> quantitatively the quality of the image. This may seem a surprising, or at >> least somewhat bleak, assessment. But consider this: In the 20 years or >> more since introduction of laser scanning confocal microscopes it has not >> been feasible, from published data, to assess how these systems compare (in >> terms of absolute units associated with measurements). No one can assert >> with confidence that instrument A in use in 1990 has better or worse >> sensitivity than instrument B operating in 2006. There is, therefore, a >> paramount need for standardized test samples and procedures for their use." >>> >>> -taken from: >>> Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization >> of fluorescence measurements from the instrument manufacturer's view >> standardization and quality assurance in fluorescence measurements ii. >> 2008. 6: p. 3-24. >>> >>> I find the second last statement above quite frustrating, especially >> from the point of view of someone who works in the industry. I can tell you >> that the experience of demoing a microscope system to a potential customer >> often goes something like this: The potential customer prepares a typical >> biological specimen that they are familiar with from experience and then >> images of this specimen are captured with the demo system. These images are >> then compared to a similar set of images that were produced on another >> competing instrument (maybe - sometimes comparison images are not >> available). Usually there are statements made to the effect of "yes/no, the >> images on this instrument look better/worse, therefore I should buy/decline >> instrument A/B". I think anyone with a good imagination can figure out that >> this is a very subjective way to compare instrument performance and that >> there are a lot of potential pitfalls involved that might bias the >> assessment one way or another. I'm not saying this isn't a good method to >> judge an microscope's general imaging performance. Indeed, you should >> always take a prospective microscope system out for a "test drive", but a >> more quantitative test that doesn't solely depend on the opinion of how >> "good" an image looks would be better for everyone in my opinion. >>> >>> And Kurt, supposing a good set of text specimens and protocols were >> developed - how could we all come to agree on them? Who would set the >> standards and govern them for microscopy? Surely the industry players would >> have much to say (or dispute!) about that. Again, as stated above, there is >> a paramount need in microscopy for standardized test samples and >> procedures. It's a cause I certainly would be willing to devote time to >> were there an organized and concerted effort to make it happen. >>> >>> Curious to hear other people's thoughts on this. >>> >>> >>> John Oreopoulos >>> Staff Scientist >>> Spectral Applied Research Inc. >>> A Division of Andor Technology >>> Richmond Hill, Ontario >>> Canada >>> www.spectral.ca >>> >>> >>> >>> On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: >>> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> hi all, >>> anyone who has worked in clinical or pre-clinical imaging will be >> familiar with the National Electrical Manufacturer's Association, or NEMA. >>> they are like a north american DIN, in that they set standards for the >> specification and performance of electrical equipment ranging from conduit >> to clinical PET scanners. >>> see here< >> http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an >> interesting list of standards. >>> recently i've been involved in using NEMA protocol NU4-2008 to >> characterise a pre-clinical PET scanner. >>> among other things, the protocol specifies the samples and methods to be >> used for quantification of the instrument sensitivity, resolution, and >> image quality. >>> my naive question for a friday afternoon is this: why dont we have a >> NEMA standard for confocal microscopes? >>> is anyone aware of this issue ever having been discussed by NEMA? >>> cheers >>> kurt >>> . >>> Prof. Kurt I. Anderson >>> Tumor Cell Migration Lab and >>> Beatson Advanced Imaging Resource (BAIR) >>> The Beatson Institute for Cancer Research >>> Garscube Estate, Switchback Road, Glasgow G61 1BD >>> * Direct Line +44 (0) 141 330 2864 >>> * Fax +44 (0) 141 942 6521 >>> [hidden email]<mailto:[hidden email] >>> <mailto:[hidden email]> >>> A company limited by guarantee; Registered in Scotland No 84170; A >> Registered Scottish Charity No SC006106 >>> The information contained in this communication may be privileged and >> confidential and is intended for the exclusive use of the addressee >> designated above. If you are not the addressee, any disclosure, >> reproduction, distribution or other dissemination or use of this >> communication is strictly prohibited. If you have received this >> transmission in error please would you be kind enough to inform us. >>> >>> >>> ________________________________ >>> This message is intended for the use of the person(s) to whom it may be >> addressed. It may contain information that is privileged, confidential, or >> otherwise protected from disclosure under applicable law. If you are not >> the intended recipient, any dissemination, distribution, copying, or use of >> this information is prohibited. If you have received this message in error, >> please permanently delete it and immediately notify the sender. Thank you. >> |
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In reply to this post by Robin Battye-2
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** That sounds like a great plan Guillermo Guillermo Marqués University Imaging Centers University of Minnesota Jackson Hall 1-151 321 Church St SE Minneapolis, MN55455 On Mar 16, 2014, at 4:58 PM, Robin Battye <[hidden email]> wrote: > If one of these calibration slides was made available for a so called road show would the community be willing to collect measurements and then post them to a public web page or forum? The slide could then move around the world from facility to facility with the goal of collecting demographics from different techniques and instruments. > > > > > Robin Battye, M.Sc., Ph. D. > Product Manager/ > Technical Sales Specialist > Mobile: (416) 302-5934 > > Spectral Applied Research > A Division of Andor Technoloy > 9078 Leslie St., Unit 11 > Richmond Hill, Ontario > L4B 3L8 > > Office: (905) 326-5040 ext. 406 > Fax: (905) 326-5041 > www.spectral.ca > > We believe that performance comes before Compromise. > > > |
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In reply to this post by Kurt Anderson-3
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hello John and Confocal Listserv Members, I would like to add to John's excellent post below. Yes, I also agree that ABRF is doing a great job on "Process and Methods" to measure optical performance and protocols for optical microscopes and confocal instruments. Regarding NEMA standards, they are the organization that is providing and managing the DICOM standards for medical imaging and in particular relevance to the microscopy world, the standards for Whole Slide Imaging ( WSI) for which I am a voting member. You can read this supplement to current DICOM standard at ftp://medical.nema.org/medical/dicom/final/sup145_ft.pdf ISO (International Standards Organization) Standards do cover microscopy and endoscopy. You can review what they do at http://www.iso.org/iso/home/standards.htm and specifically the microscopy standards at ISO TC172/SC5 http://www.iso.org/iso/home/store/catalogue_tc/catalogue_tc_browse.htm?commid=53720 The US organization for microscopy standards is called OEOSC or Optics and Electro-Optics Standards Council which is the US arm of ISO standards. You can see they do at http://www.optstd.org/index.htm and specifically for microscopes at http://www.optstd.org/isotc172.htm ISO standards ( originally DIN standards) concentrate on manufacturing standards. Currently relevant standards are being developed for: ISO/CD 19056-1 Microscopes -- Definition and measurement of illumination properties -- Part 1: Image brightness and uniformity in bright field microscopy ISO/DIS 19012-3 Microscopes -- Designation of microscope objectives -- Part 3: Spectral transmittance ISO/CD 18221 Microscopes -- Microscopes with digital imaging displays -- Information provided to the user regarding imaging performance Currently there are 37 published standards and 11 more in preparation. I am the US delegation leader for ISO standards in microscopy. If you have additional questions you can contact me off the listserv at [hidden email] Respectfully submitted, Stan Schwartz Nikon Instruments Inc. Melville, NY 11747 USA On Mon, 17 Mar 2014 09:20:00 -0400, John Oreopoulos <[hidden email]> wrote: >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >Yes, I would agree that the ABRF has made the greatest efforts in trying to measure microscope imaging performance on a somewhat large scale and develop imaging diagnostic protocols/standards. Their two recent publications are evidence of that: > >Stack, R. F., Bayles, C. J., Girard, A. M., Martin, K., Opansky, C., Schulz, K., et al. (2011). Quality assurance testing for modern optical imaging systems. Microscopy and Microanalysis, 17, 598-606. > >Cole, R. W., Thibault, M., Bayles, C. J., Eason, B., Girard, A. M., Jinadasa, T., et al. (2013). International test results for objective lens quality, resolution, spectral accuracy and spectral separation for confocal laser scanning microscopes. Microscopy and Microanalysis, 19, 1653-1668. > >They would be a logical organization to figure this sort of thing out. I suppose the RMS or a similar organization would also be up to the task and have the right resources for this. The acceptance of any kind of imaging standard (for example, see the USAF resolution target: http://en.wikipedia.org/wiki/1951_USAF_resolution_test_chart) would require some level of "metrological traceability" or validation as is described by NIST: > >http://www.nist.gov/traceability/nist_traceability_policy_external.cfm > >Again, it's not quite clear to me who the NEMA equivalent of the microscopy world is or if even such a body exists. What's interesting about NEMA is that its main members are big electrical companies like GE, 3M, and Siemans, so clearly there was an effort on the electrical industry's part to establish these electrical standards: > >http://www.nema.org/About/History/Pages/Model-of-Industrial-Self-Government.aspx > >I think ISO is the largest standardization organization I'm aware of: > >http://en.wikipedia.org/wiki/International_Organization_for_Standardization > >Are there any fluorescence microscopy or imaging ISO standards? > > > >John Oreopoulos >Staff Scientist >Spectral Applied Research Inc. >A Division of Andor Technology >Richmond Hill, Ontario >Canada >www.spectral.ca > > > >On 2014-03-17, at 7:00 AM, phil laissue wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> But isn't that exactly what the ABRF is trying to establish? >> >> http://www.ncbi.nlm.nih.gov/pubmed/21477410 >> >> >> >> _____________________________________ >> Philippe Laissue, PhD, Bioimaging Manager >> School of Biological Sciences, Room 4.17 >> University of Essex, Colchester CO4 3SQ, UK >> (0044) 01206 872246 / (0044) 07842 676 456 >> [hidden email] >> privatewww.essex.ac.uk/~plaissue >> >> >> On 16 March 2014 03:43, John Oreopoulos <[hidden email]> wrote: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> Well, that's very interesting. This is exactly the kind of test specimen I >>> had in mind for fluorescence microscopy. I had not heard of this company >>> before now. Thank you Glenn for pointing them out. I think I'll have to try >>> using one of their test slides. What I find most interesting here is >>> Argolight's ability to manufacture complex 3D structures with the >>> fluorescent brush technology. The question then becomes - what's a >>> good/challenging test specimen 3D pattern for confocal microscopy that >>> everyone could all agree on? >>> >>> John Oreopoulos >>> Staff Scientist >>> Spectral Applied Research Inc. >>> A Division of Andor Technology >>> Richmond Hill, Ontario >>> Canada >>> www.spectral.ca >>> >>> >>> On 2014-03-15, at 4:56 PM, Glenn Merrill-Skoloff wrote: >>> >>>> ***** >>>> To join, leave or search the confocal microscopy listserv, go to: >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>>> ***** >>>> >>>> There's a commercially available product that claims to meet most of the >>> requirements John lists, below. Has anyone used this slide? >>>> >>>> http://argolight.com/product-micro/ >>>> >>>> My Best, >>>> -- Glenn >>>> >>>> Glenn Merrill-Skoloff >>>> Division of Hemostasis and Thrombosis >>>> Director, Intravital Microscopy Core >>>> >>>> 617-735-4040 (Office) >>>> 617-735-4007 (Lab) >>>> 617-735-4000 (Fax) >>>> >>>> >>>> >>>> From: John Oreopoulos <[hidden email]<mailto: >>> [hidden email]>> >>>> Reply-To: Confocal Microscopy List <[hidden email] >>> <mailto:[hidden email]>> >>>> Date: Saturday, March 15, 2014 at 12:35 AM >>>> To: "[hidden email]<mailto: >>> [hidden email]>" <[hidden email] >>> <mailto:[hidden email]>> >>>> Subject: Re: Confocal NEMA? >>>> >>>> ***** >>>> To join, leave or search the confocal microscopy listserv, go to: >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>>> ***** >>>> >>>> Kurt, >>>> >>>> I'm not familiar with NEMA, but the general topic of >>> testing/standardizing confocal sensitivity, resolution, and image quality >>> is of great interest to me. If you search the confocal listserver archive, >>> you'll find several postings about this. In theory, you could develop a >>> standard test for confocal sensitivity, resolution, and image quality, but >>> what should the test specimen be? Others have worked on this to some >>> extent, and a few names that come to mind are Fred Brakenhoff, Claire >>> Brown, Robert Zucker, and Mike Model. I recommend looking up their papers >>> to see what sorts of test specimens have been proposed and tried in the >>> past. Unfortunately, I don't think there is one single test specimen that >>> can be used to determine all instrument parameters of interest >>> (sensitivity, resolution - spatial, temporal, spectral-, and image quality, >>> etc.). >>>> >>>> In my mind, an ideal test specimen for confocal fluorescence imaging >>> would have the following properties: >>>> >>>> 1. Be cheap, readily available, and reproducible to a high degree >>>> 2. Able to absorb and emit light over broad range of wavelengths >>>> 3. Be portable and emit constant intensity (no photobleaching) perhaps >>> even with a known number of photons under certain conditions >>>> 4. Does not saturate easily, and has a linear response to excitation >>> light power >>>> 5. Has a known 3D structure of specific shapes/patterns/sizes with a >>> high level of precision determined by some other imaging technique >>> (electron microscopy for example). >>>> 6. Has a refractive index close to water or glass >>>> >>>> Again, I don't think such a sample exists, but if you can think of one, >>> let me know! I think one of the other problems here is: how would you >>> define and quantify things like "image quality"? I'm curious how NEMA does >>> this for PET. Spatial resolution is a bit more straightforward to quantify, >>> although here too there are different ways to define resolution and ways to >>> measure it. Sensitivity measurements are complicated by the need to keep >>> many other imaging parameters constant when examining day-to-day >>> performance/sensitivity of one instrument, or comparing the performance of >>> two different instruments - see Jim Pawley's "39 steps". >>>> >>>> Your question also brings to mind a quote from another very well-written >>> article on the topic which I never forget: >>>> >>>> "This complex relationship between qualitative and quantitative content >>> of fluorescence images also expresses itself in the way that commercial >>> instruments are assessed. On the one hand it is unavoidable that users >>> initially are strongly influenced by the visual quality of the images >>> presented by the instrument. On the other hand the longer term scientific >>> value of the instrument depends crucially both on the quality of the visual >>> information and the effectiveness with which it can be reduced for >>> quantitative analysis... The increased complexity of a fluorescent image >>> (in terms of the number of dimensions that can be measured), and the >>> variability of sample preparation, makes it less easy to assess >>> quantitatively the quality of the image. This may seem a surprising, or at >>> least somewhat bleak, assessment. But consider this: In the 20 years or >>> more since introduction of laser scanning confocal microscopes it has not >>> been feasible, from published data, to assess how these systems compare (in >>> terms of absolute units associated with measurements). No one can assert >>> with confidence that instrument A in use in 1990 has better or worse >>> sensitivity than instrument B operating in 2006. There is, therefore, a >>> paramount need for standardized test samples and procedures for their use." >>>> >>>> -taken from: >>>> Dixon, A., T. Heinlein, and R. Wolleschensky, Need for standardization >>> of fluorescence measurements from the instrument manufacturer's view >>> standardization and quality assurance in fluorescence measurements ii. >>> 2008. 6: p. 3-24. >>>> >>>> I find the second last statement above quite frustrating, especially >>> from the point of view of someone who works in the industry. I can tell you >>> that the experience of demoing a microscope system to a potential customer >>> often goes something like this: The potential customer prepares a typical >>> biological specimen that they are familiar with from experience and then >>> images of this specimen are captured with the demo system. These images are >>> then compared to a similar set of images that were produced on another >>> competing instrument (maybe - sometimes comparison images are not >>> available). Usually there are statements made to the effect of "yes/no, the >>> images on this instrument look better/worse, therefore I should buy/decline >>> instrument A/B". I think anyone with a good imagination can figure out that >>> this is a very subjective way to compare instrument performance and that >>> there are a lot of potential pitfalls involved that might bias the >>> assessment one way or another. I'm not saying this isn't a good method to >>> judge an microscope's general imaging performance. Indeed, you should >>> always take a prospective microscope system out for a "test drive", but a >>> more quantitative test that doesn't solely depend on the opinion of how >>> "good" an image looks would be better for everyone in my opinion. >>>> >>>> And Kurt, supposing a good set of text specimens and protocols were >>> developed - how could we all come to agree on them? Who would set the >>> standards and govern them for microscopy? Surely the industry players would >>> have much to say (or dispute!) about that. Again, as stated above, there is >>> a paramount need in microscopy for standardized test samples and >>> procedures. It's a cause I certainly would be willing to devote time to >>> were there an organized and concerted effort to make it happen. >>>> >>>> Curious to hear other people's thoughts on this. >>>> >>>> >>>> John Oreopoulos >>>> Staff Scientist >>>> Spectral Applied Research Inc. >>>> A Division of Andor Technology >>>> Richmond Hill, Ontario >>>> Canada >>>> www.spectral.ca >>>> >>>> >>>> >>>> On 2014-03-14, at 1:14 PM, Kurt Anderson wrote: >>>> >>>> ***** >>>> To join, leave or search the confocal microscopy listserv, go to: >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>>> ***** >>>> hi all, >>>> anyone who has worked in clinical or pre-clinical imaging will be >>> familiar with the National Electrical Manufacturer's Association, or NEMA. >>>> they are like a north american DIN, in that they set standards for the >>> specification and performance of electrical equipment ranging from conduit >>> to clinical PET scanners. >>>> see here< >>> http://www.nema.org/Standards/Pages/All-Standards-by-Product.aspx> for an >>> interesting list of standards. >>>> recently i've been involved in using NEMA protocol NU4-2008 to >>> characterise a pre-clinical PET scanner. >>>> among other things, the protocol specifies the samples and methods to be >>> used for quantification of the instrument sensitivity, resolution, and >>> image quality. >>>> my naive question for a friday afternoon is this: why dont we have a >>> NEMA standard for confocal microscopes? >>>> is anyone aware of this issue ever having been discussed by NEMA? >>>> cheers >>>> kurt >>>> . >>>> Prof. Kurt I. Anderson >>>> Tumor Cell Migration Lab and >>>> Beatson Advanced Imaging Resource (BAIR) >>>> The Beatson Institute for Cancer Research >>>> Garscube Estate, Switchback Road, Glasgow G61 1BD >>>> * Direct Line +44 (0) 141 330 2864 >>>> * Fax +44 (0) 141 942 6521 >>>> [hidden email]<mailto:[hidden email] >>>> <mailto:[hidden email]> >>>> A company limited by guarantee; Registered in Scotland No 84170; A >>> Registered Scottish Charity No SC006106 >>>> The information contained in this communication may be privileged and >>> confidential and is intended for the exclusive use of the addressee >>> designated above. If you are not the addressee, any disclosure, >>> reproduction, distribution or other dissemination or use of this >>> communication is strictly prohibited. If you have received this >>> transmission in error please would you be kind enough to inform us. >>>> >>>> >>>> ________________________________ >>>> This message is intended for the use of the person(s) to whom it may be >>> addressed. It may contain information that is privileged, confidential, or >>> otherwise protected from disclosure under applicable law. If you are not >>> the intended recipient, any dissemination, distribution, copying, or use of >>> this information is prohibited. If you have received this message in error, >>> please permanently delete it and immediately notify the sender. Thank you. >>> |
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In reply to this post by Kurt Anderson-3
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I want to thank John for pointing out ABRF's (http://www.abrf.org/index.cfm/group.show/LightMicroscopyResearchGroup.54.ht m) involvement in this very process. This is exactly what the Light Microscopy research group, with the help of imagers world-wide, is trying to establish. We have distributed free test samples as well as detailed protocols in two previous studies, which tested a wide variety of instrument parameters. John has provided the references for those two studies, check them out for more details including protocols and world-wide results. For a more in-depth look at PSF see: Cole, R.W., Jinadasa, T., and Brown, C.M. (2011) Resolution and Quality Control of Confocal Microscopy Optics. Nature Prot. 6 (12): 1929-1941. doi:10.1038/nprot.2011.407 The current study, about to go public, will move away from the coverslip and provide data and metrics from where we all image, i.e. 0-100um above the coverslip. We have spent over a year brain-storming, designing and testing a sample that will work and believe that we have achieved what we set out to accomplish. I don't not believe that there is a single test specimen that will work for all the necessary tests. The goal was and still is, to provide very inexpensive test specimens and protocols along with expectable metrics to enable users to routinely check their instruments. Watch the list sever for more information regarding the upcoming test and once again free samples and protocols. Thanks to all that have participated in previous studies! Richard Cole Research Scientist V Director: Advanced Light Microscopy & Image Analysis Core Wadsworth Center Research Assistant Professor Dept. of Biomedical Sciences School of Public Health State University of New York P.O. Box 509 Albany N.Y. 12201-0509 518-474-7048 Phone 518-474-4430 Fax Email <mailto:[hidden email]> [hidden email] Website <http://www.wadsworth.org/cores/alm/index.htm> www.wadsworth.org/cores/alm/index.htm |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Richard, I'm very curious to find out what your third ABRF study will be using as a "thick", cheap test specimen for confocal imaging. How does it compare to the precision of dimensions for the test specimen listed on the Argolight webpage (one of the Argolight patterns is a cylinder with about 250 nm thick fluorescent walls extending 30 um deep, 50 nm precision of features). What shape/pattern will your test specimen contain? John Oreopoulos Staff Scientist Spectral Applied Research Inc. A Division of Andor Technology Richmond Hill, Ontario Canada www.spectral.ca On 2014-03-18, at 8:46 AM, Richard Cole wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > I want to thank John for pointing out ABRF's > (http://www.abrf.org/index.cfm/group.show/LightMicroscopyResearchGroup.54.ht > m) involvement in this very process. This is exactly what the Light > Microscopy research group, with the help of imagers world-wide, is trying to > establish. We have distributed free test samples as well as detailed > protocols in two previous studies, which tested a wide variety of instrument > parameters. John has provided the references for those two studies, check > them out for more details including protocols and world-wide results. For a > more in-depth look at PSF see: Cole, R.W., Jinadasa, T., and Brown, C.M. > (2011) Resolution and Quality Control of Confocal Microscopy Optics. Nature > Prot. 6 (12): 1929-1941. doi:10.1038/nprot.2011.407 > > > > The current study, about to go public, will move away from the coverslip and > provide data and metrics from where we all image, i.e. 0-100um above the > coverslip. We have spent over a year brain-storming, designing and testing > a sample that will work and believe that we have achieved what we set out to > accomplish. I don't not believe that there is a single test specimen that > will work for all the necessary tests. The goal was and still is, to provide > very inexpensive test specimens and protocols along with expectable metrics > to enable users to routinely check their instruments. > > > > Watch the list sever for more information regarding the upcoming test and > once again free samples and protocols. > > > > Thanks to all that have participated in previous studies! > > > > > > > > Richard Cole > Research Scientist V > Director: Advanced Light Microscopy & Image Analysis Core > Wadsworth Center > > > > Research Assistant Professor > Dept. of Biomedical Sciences > School of Public Health State University of New York > > > P.O. Box 509 Albany N.Y. 12201-0509 > 518-474-7048 Phone > 518-474-4430 Fax > > > > Email <mailto:[hidden email]> [hidden email] > > Website <http://www.wadsworth.org/cores/alm/index.htm> > www.wadsworth.org/cores/alm/index.htm > > > > > > |
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In reply to this post by Kurt Anderson-3
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** There is a paper by Khan and Brumberg "An inexpensive fluorescent graticule" in Microscopy Today, vol 22, 26-27 (2014). Mike Model |
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