Re: Confocal calibration & performance assesment **vendor response**

[Kilgore, Jason A.]

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** Vendor response **

Hi, Javier,

For a uniform fluorescence standard, I recommend the Fluor-Ref slides.  These are sold or re-sold by various places and are fairly inexpensive.  These are plastic slides that are autofluorescent in four different wavelengths and are incredibly uniform and photostable.  The drawbacks are that they are extremely bright, so much so that you often either have to throw in neutral density filters or image an off-wavelength slide.  Also, they are thicker than a standard slide.  Here's the webpage from Ted Pella:  http://www.tedpella.com/histo_html/fluor.htm.aspx or resold as a set of four from Fisher Scientific: https://www.fishersci.com/shop/products/NC0158348/nc0158348#?keyword=FLUOR-REF+SET+OF+4+SLIDES 

There are other published methods for uniformity slides (such as a simple solution of fluorescein under a coverslip, or putting dye in agar) or labeling coated slides, but nothing comes close to the Fluor-Ref slides for photostability, storage stability, and uniformity of intensity.

There is a good and fairly inexpensive option for some of the other needs you mention:  FocalCheck microspheres slides.  There are different slides, but probably Slide #1 is best for your needs, catalog F36909.  The top row has options where you align rings of different colors to check alignment, or rings and cores of the microspheres. There are also TetraSpeck beads of three different sizes (4, 1, and 0.5 um) that are broadly fluorescent, which you can use for PSF as well as alignment.  The bottom row has 6um beads of five different relative intensities, for doing intensity calibrations.  
Webpage from Thermo Fisher Scientific:  https://www.thermofisher.com/order/catalog/product/F36909
Manual: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/mp36909.pdf 

We sell fluorescent beads of smaller sizes, too, if you need them (such as 20 nm) of different wavelengths, for PSF measures.

I hope this helps, but please feel free to contact me offline if you wish more info (such as a PowerPoint I have showing images from that last slide).

Jason


Jason A. Kilgore
Technical Application Scientist
Molecular Probes / EVOS Tech Support
Thermo Fisher Scientific
 
1-800-955-6288 then option 4, then option 3, then option 2.
Or dial direct at [phone]
[hidden email]
 
This communication is intended solely for the individual/entity to whom it is addressed. It may contain confidential or legally privileged information.  Any unauthorized disclosure or copying is prohibited and may be unlawful. If you have received this communication in error, please notify the sender immediately and delete it from your system.
 
 


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Fco. Javier Díez Guerra
Sent: Wednesday, August 30, 2017 5:10 AM
To: [hidden email]
Subject: Confocal calibration & performance assesment

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Dear confocalists,

I would like to know about the preferred methods used by facility managers to regularly asses the performance of their confocal (and
wide-field) microscopes. In other words, which are the most used and reliable methods to check different features of confocal and wide-field microscope peformance?.

Particularly, we'd like to check:

- illumination uniformity across FOV, now we use a slide coated with fluorescent secondary antibody.

- XYZ Chromatic aberration (exciting 405, 488, 561 and 640 nm), now we use 1um beads that can be excited with all laser lines.

- XYZ Resolution, we use smaller beads (0,17um) to build and analyze PSFs (for each wavelength)

- Laser power and stability: for power, we use a power meter from Newport placed at the objective exit, for stability, we capture long time-series in reflexion mode with an empty preparation.

- We do not know how to check detector (PMT, GAsP, Hybrid, etc) sensitivity and SNR.

Thanks for your input.


--
Fco. Javier Diez-Guerra, PhD

Profesor Titular UAM
Servicio de Microscopía Confocal
Centro de Biologia Molecular Severo Ochoa C/ Nicolás Cabrera, 1 Campus de Cantoblanco
28049 Madrid
SPAIN

Tel     [phone]
e-mail: [hidden email]

[George McNamara]

*****
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Dear listserv,

Fluor-Ref slides ... pick up a box or case from Chroma at a conference.

As for Thermo: how about giving away THIN fluorescent plastic specimens
.. can include your logo (and no, it would not fall under the category
of pens or coffee cups that you can no longer give out to U.S. academic
medical center staff).

Back to Chroma slides ... if not going to 'Neuo' or 'Cell Bio', and
cannot wait for M&M 8/2018 in Baltimore, as an alternative you can use a
brightly colored clipboard(s). Come ot think of it, Thermo could give
out branded clipboards as big reference slides.

enjoy,

George


On 9/1/2017 1:26 PM, Kilgore, Jason A. wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
>
> ** Vendor response **
>
> Hi, Javier,
>
> For a uniform fluorescence standard, I recommend the Fluor-Ref slides.  These are sold or re-sold by various places and are fairly inexpensive.  These are plastic slides that are autofluorescent in four different wavelengths and are incredibly uniform and photostable.  The drawbacks are that they are extremely bright, so much so that you often either have to throw in neutral density filters or image an off-wavelength slide.  Also, they are thicker than a standard slide.  Here's the webpage from Ted Pella:  http://www.tedpella.com/histo_html/fluor.htm.aspx or resold as a set of four from Fisher Scientific: https://www.fishersci.com/shop/products/NC0158348/nc0158348#?keyword=FLUOR-REF+SET+OF+4+SLIDES
>
> There are other published methods for uniformity slides (such as a simple solution of fluorescein under a coverslip, or putting dye in agar) or labeling coated slides, but nothing comes close to the Fluor-Ref slides for photostability, storage stability, and uniformity of intensity.
>
> There is a good and fairly inexpensive option for some of the other needs you mention:  FocalCheck microspheres slides.  There are different slides, but probably Slide #1 is best for your needs, catalog F36909.  The top row has options where you align rings of different colors to check alignment, or rings and cores of the microspheres. There are also TetraSpeck beads of three different sizes (4, 1, and 0.5 um) that are broadly fluorescent, which you can use for PSF as well as alignment.  The bottom row has 6um beads of five different relative intensities, for doing intensity calibrations.
> Webpage from Thermo Fisher Scientific:  https://www.thermofisher.com/order/catalog/product/F36909
> Manual: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/mp36909.pdf
>
> We sell fluorescent beads of smaller sizes, too, if you need them (such as 20 nm) of different wavelengths, for PSF measures.
>
> I hope this helps, but please feel free to contact me offline if you wish more info (such as a PowerPoint I have showing images from that last slide).
>
> Jason
>
>
> Jason A. Kilgore
> Technical Application Scientist
> Molecular Probes / EVOS Tech Support
> Thermo Fisher Scientific
>  
> 1-800-955-6288 then option 4, then option 3, then option 2.
> Or dial direct at [phone]
> [hidden email]
>  
> This communication is intended solely for the individual/entity to whom it is addressed. It may contain confidential or legally privileged information.  Any unauthorized disclosure or copying is prohibited and may be unlawful. If you have received this communication in error, please notify the sender immediately and delete it from your system.
>  
>  
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Fco. Javier Díez Guerra
> Sent: Wednesday, August 30, 2017 5:10 AM
> To: [hidden email]
> Subject: Confocal calibration & performance assesment
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear confocalists,
>
> I would like to know about the preferred methods used by facility managers to regularly asses the performance of their confocal (and
> wide-field) microscopes. In other words, which are the most used and reliable methods to check different features of confocal and wide-field microscope peformance?.
>
> Particularly, we'd like to check:
>
> - illumination uniformity across FOV, now we use a slide coated with fluorescent secondary antibody.
>
> - XYZ Chromatic aberration (exciting 405, 488, 561 and 640 nm), now we use 1um beads that can be excited with all laser lines.
>
> - XYZ Resolution, we use smaller beads (0,17um) to build and analyze PSFs (for each wavelength)
>
> - Laser power and stability: for power, we use a power meter from Newport placed at the objective exit, for stability, we capture long time-series in reflexion mode with an empty preparation.
>
> - We do not know how to check detector (PMT, GAsP, Hybrid, etc) sensitivity and SNR.
>
> Thanks for your input.
>
>
> --
> Fco. Javier Diez-Guerra, PhD
>
> Profesor Titular UAM
> Servicio de Microscopía Confocal
> Centro de Biologia Molecular Severo Ochoa C/ Nicolás Cabrera, 1 Campus de Cantoblanco
> 28049 Madrid
> SPAIN
>
> Tel     [phone]
> e-mail: [hidden email]

--


George McNamara, PhD
Baltimore, MD 21231
[hidden email]
https://www.linkedin.com/in/georgemcnamara
https://works.bepress.com/gmcnamara/75   (may need to use Microsoft Edge or Firefox, rather than Google Chrome)
http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650
http://confocal.jhu.edu

July 2017 Current Protocols article, open access:
UNIT 4.4 Microscopy and Image Analysis
http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/abstract
supporting materials direct link is
http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/full#hg0404-sec-0023
figures at
http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/figures

[mmodel]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

Chroma slides may be too bright for wide-field (though, to tell the truth, I don't see any need to monitor the performance of lamp-based microscopes), I don't know about Molecular Probes' slides. But one can sometimes use Chroma slides in an "unintended" way, something like a red slide with blue illumination, and then the brightness becomes more reasonable.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of George McNamara
Sent: Saturday, September 2, 2017 1:55 PM
To: [hidden email]
Subject: Fluor-Ref slides ... pick up a box or case from Chroma at a conference ... Re: Confocal calibration & performance assesment **vendor response**

*****
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Post images on https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=fimjaHIaIMnlqdqyWYm19D0CvjyCBrB08m8x17FX6dk%3D&reserved=0 and include the link in your posting.
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Dear listserv,

Fluor-Ref slides ... pick up a box or case from Chroma at a conference.

As for Thermo: how about giving away THIN fluorescent plastic specimens .. can include your logo (and no, it would not fall under the category of pens or coffee cups that you can no longer give out to U.S. academic medical center staff).

Back to Chroma slides ... if not going to 'Neuo' or 'Cell Bio', and cannot wait for M&M 8/2018 in Baltimore, as an alternative you can use a brightly colored clipboard(s). Come ot think of it, Thermo could give out branded clipboards as big reference slides.

enjoy,

George


On 9/1/2017 1:26 PM, Kilgore, Jason A. wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
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> umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cmmodel
> %40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e
> 7dd15f26134%7C1&sdata=n6OeQLr2NtHtB320h62kEiK%2BZZJI5PuqSWoqcefDnME%3D
> &reserved=0 Post images on
> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=fimjaHIaIMnlqdqyWYm19D0CvjyCBrB08m8x17FX6dk%3D&reserved=0 and include the link in your posting.
> *****
>
>
> ** Vendor response **
>
> Hi, Javier,
>
> For a uniform fluorescence standard, I recommend the Fluor-Ref slides.  
> These are sold or re-sold by various places and are fairly
> inexpensive.  These are plastic slides that are autofluorescent in
> four different wavelengths and are incredibly uniform and photostable.  
> The drawbacks are that they are extremely bright, so much so that you
> often either have to throw in neutral density filters or image an
> off-wavelength slide.  Also, they are thicker than a standard slide.  
> Here's the webpage from Ted Pella:  
> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.te
> dpella.com%2Fhisto_html%2Ffluor.htm.aspx&data=01%7C01%7Cmmodel%40KENT.
> EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26
> 134%7C1&sdata=BcsiKMcDnxqoK1jIMqN2mZHgKzYkaN%2BzWhCmiq3iyCA%3D&reserve
> d=0 or resold as a set of four from Fisher Scientific:
> https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.f
> ishersci.com%2Fshop%2Fproducts%2FNC0158348%2Fnc0158348%23%3Fkeyword%3D
> FLUOR-REF%2BSET%2BOF%2B4%2BSLIDES&data=01%7C01%7Cmmodel%40KENT.EDU%7C3
> 6cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1
> &sdata=xv9a4R6u7RpajF1wKTSSuvTx5cB71pgNea6XQrt8RKg%3D&reserved=0
>
> There are other published methods for uniformity slides (such as a simple solution of fluorescein under a coverslip, or putting dye in agar) or labeling coated slides, but nothing comes close to the Fluor-Ref slides for photostability, storage stability, and uniformity of intensity.
>
> There is a good and fairly inexpensive option for some of the other needs you mention:  FocalCheck microspheres slides.  There are different slides, but probably Slide #1 is best for your needs, catalog F36909.  The top row has options where you align rings of different colors to check alignment, or rings and cores of the microspheres. There are also TetraSpeck beads of three different sizes (4, 1, and 0.5 um) that are broadly fluorescent, which you can use for PSF as well as alignment.  The bottom row has 6um beads of five different relative intensities, for doing intensity calibrations.
> Webpage from Thermo Fisher Scientific:  
> https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.t
> hermofisher.com%2Forder%2Fcatalog%2Fproduct%2FF36909&data=01%7C01%7Cmm
> odel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018
> f73e7dd15f26134%7C1&sdata=IbU37oBjbMd9zZE1RKzb4Lp8g6TWlZjE3FJAc6lxtJ0%
> 3D&reserved=0
> Manual:
> https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fasset
> s.thermofisher.com%2FTFS-Assets%2FLSG%2Fmanuals%2Fmp36909.pdf&data=01%
> 7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a
> 1ec44d018f73e7dd15f26134%7C1&sdata=rBOAKsVfx3HvTl0Y2kif8s9lVGzwfgfHYZc
> %2BYsl1mY0%3D&reserved=0
>
> We sell fluorescent beads of smaller sizes, too, if you need them (such as 20 nm) of different wavelengths, for PSF measures.
>
> I hope this helps, but please feel free to contact me offline if you wish more info (such as a PowerPoint I have showing images from that last slide).
>
> Jason
>
>
> Jason A. Kilgore
> Technical Application Scientist
> Molecular Probes / EVOS Tech Support
> Thermo Fisher Scientific
>  
> 1-800-955-6288 then option 4, then option 3, then option 2.
> Or dial direct at [phone]
> [hidden email]
>  
> This communication is intended solely for the individual/entity to whom it is addressed. It may contain confidential or legally privileged information.  Any unauthorized disclosure or copying is prohibited and may be unlawful. If you have received this communication in error, please notify the sender immediately and delete it from your system.
>  
>  
>
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]] On Behalf Of Fco. Javier
> Díez Guerra
> Sent: Wednesday, August 30, 2017 5:10 AM
> To: [hidden email]
> Subject: Confocal calibration & performance assesment
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.
> umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cmmodel
> %40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e
> 7dd15f26134%7C1&sdata=n6OeQLr2NtHtB320h62kEiK%2BZZJI5PuqSWoqcefDnME%3D
> &reserved=0 Post images on
> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=fimjaHIaIMnlqdqyWYm19D0CvjyCBrB08m8x17FX6dk%3D&reserved=0 and include the link in your posting.
> *****
>
> Dear confocalists,
>
> I would like to know about the preferred methods used by facility
> managers to regularly asses the performance of their confocal (and
> wide-field) microscopes. In other words, which are the most used and reliable methods to check different features of confocal and wide-field microscope peformance?.
>
> Particularly, we'd like to check:
>
> - illumination uniformity across FOV, now we use a slide coated with fluorescent secondary antibody.
>
> - XYZ Chromatic aberration (exciting 405, 488, 561 and 640 nm), now we use 1um beads that can be excited with all laser lines.
>
> - XYZ Resolution, we use smaller beads (0,17um) to build and analyze
> PSFs (for each wavelength)
>
> - Laser power and stability: for power, we use a power meter from Newport placed at the objective exit, for stability, we capture long time-series in reflexion mode with an empty preparation.
>
> - We do not know how to check detector (PMT, GAsP, Hybrid, etc) sensitivity and SNR.
>
> Thanks for your input.
>
>
> --
> Fco. Javier Diez-Guerra, PhD
>
> Profesor Titular UAM
> Servicio de Microscopía Confocal
> Centro de Biologia Molecular Severo Ochoa C/ Nicolás Cabrera, 1 Campus
> de Cantoblanco
> 28049 Madrid
> SPAIN
>
> Tel     [phone]
> e-mail: [hidden email]

--


George McNamara, PhD
Baltimore, MD 21231
[hidden email]
https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.linkedin.com%2Fin%2Fgeorgemcnamara&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=rU0v72Nc%2FLwoUg%2FhXnLPwRjmdYi2%2B9ZxzKWvtJb0kZQ%3D&reserved=0
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https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fmyncbi%2Fbrowse%2Fcollection%2F44962650&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=TaZvjTkgLYCXqeFztr4ziLPLnQWZ0JP2m45tX7A6NHg%3D&reserved=0
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July 2017 Current Protocols article, open access:
UNIT 4.4 Microscopy and Image Analysis
https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fonlinelibrary.wiley.com%2Fdoi%2F10.1002%2Fcphg.42%2Fabstract&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=91n47cpV04FX838CK9nuoYazFG6t7eIJ6hJ0ZG5ml1s%3D&reserved=0
supporting materials direct link is
https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fonlinelibrary.wiley.com%2Fdoi%2F10.1002%2Fcphg.42%2Ffull%23hg0404-sec-0023&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=rSz%2FvrvisPznVbRIHDvfUFiUCCXDgpv9rFEccjtrkd4%3D&reserved=0
figures at
https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fonlinelibrary.wiley.com%2Fdoi%2F10.1002%2Fcphg.42%2Ffigures&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=Tw2nqrqlQbOBWbTaAvxy6i4vXvwytQ8Ek3a5Zhz3IDI%3D&reserved=0

[George McNamara]

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*****

the challenge with plastic slides is the thickness is a lot thicker than
most specimens, so shading is not quite the same (advantage: confocal!).
This is where Michael's "inner filter effect" high concentration of
fluorophore has a big advantage over plastic.

Brightness: LED epi-illuminator power can be decreased. ND filters can
be put in excitation path for marc lamps and metal halide lamps. sCMOS
(and some CCDs) can be used with very short exposure time (<<1 millisecond).


On 9/2/2017 2:11 PM, MODEL, MICHAEL wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Chroma slides may be too bright for wide-field (though, to tell the truth, I don't see any need to monitor the performance of lamp-based microscopes), I don't know about Molecular Probes' slides. But one can sometimes use Chroma slides in an "unintended" way, something like a red slide with blue illumination, and then the brightness becomes more reasonable.
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of George McNamara
> Sent: Saturday, September 2, 2017 1:55 PM
> To: [hidden email]
> Subject: Fluor-Ref slides ... pick up a box or case from Chroma at a conference ... Re: Confocal calibration & performance assesment **vendor response**
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=n6OeQLr2NtHtB320h62kEiK%2BZZJI5PuqSWoqcefDnME%3D&reserved=0
> Post images on https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=fimjaHIaIMnlqdqyWYm19D0CvjyCBrB08m8x17FX6dk%3D&reserved=0 and include the link in your posting.
> *****
>
> Dear listserv,
>
> Fluor-Ref slides ... pick up a box or case from Chroma at a conference.
>
> As for Thermo: how about giving away THIN fluorescent plastic specimens .. can include your logo (and no, it would not fall under the category of pens or coffee cups that you can no longer give out to U.S. academic medical center staff).
>
> Back to Chroma slides ... if not going to 'Neuo' or 'Cell Bio', and cannot wait for M&M 8/2018 in Baltimore, as an alternative you can use a brightly colored clipboard(s). Come ot think of it, Thermo could give out branded clipboards as big reference slides.
>
> enjoy,
>
> George
>
>
> On 9/1/2017 1:26 PM, Kilgore, Jason A. wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.
>> umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cmmodel
>> %40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e
>> 7dd15f26134%7C1&sdata=n6OeQLr2NtHtB320h62kEiK%2BZZJI5PuqSWoqcefDnME%3D
>> &reserved=0 Post images on
>> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=fimjaHIaIMnlqdqyWYm19D0CvjyCBrB08m8x17FX6dk%3D&reserved=0 and include the link in your posting.
>> *****
>>
>>
>> ** Vendor response **
>>
>> Hi, Javier,
>>
>> For a uniform fluorescence standard, I recommend the Fluor-Ref slides.
>> These are sold or re-sold by various places and are fairly
>> inexpensive.  These are plastic slides that are autofluorescent in
>> four different wavelengths and are incredibly uniform and photostable.
>> The drawbacks are that they are extremely bright, so much so that you
>> often either have to throw in neutral density filters or image an
>> off-wavelength slide.  Also, they are thicker than a standard slide.
>> Here's the webpage from Ted Pella:
>> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.te
>> dpella.com%2Fhisto_html%2Ffluor.htm.aspx&data=01%7C01%7Cmmodel%40KENT.
>> EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26
>> 134%7C1&sdata=BcsiKMcDnxqoK1jIMqN2mZHgKzYkaN%2BzWhCmiq3iyCA%3D&reserve
>> d=0 or resold as a set of four from Fisher Scientific:
>> https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.f
>> ishersci.com%2Fshop%2Fproducts%2FNC0158348%2Fnc0158348%23%3Fkeyword%3D
>> FLUOR-REF%2BSET%2BOF%2B4%2BSLIDES&data=01%7C01%7Cmmodel%40KENT.EDU%7C3
>> 6cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1
>> &sdata=xv9a4R6u7RpajF1wKTSSuvTx5cB71pgNea6XQrt8RKg%3D&reserved=0
>>
>> There are other published methods for uniformity slides (such as a simple solution of fluorescein under a coverslip, or putting dye in agar) or labeling coated slides, but nothing comes close to the Fluor-Ref slides for photostability, storage stability, and uniformity of intensity.
>>
>> There is a good and fairly inexpensive option for some of the other needs you mention:  FocalCheck microspheres slides.  There are different slides, but probably Slide #1 is best for your needs, catalog F36909.  The top row has options where you align rings of different colors to check alignment, or rings and cores of the microspheres. There are also TetraSpeck beads of three different sizes (4, 1, and 0.5 um) that are broadly fluorescent, which you can use for PSF as well as alignment.  The bottom row has 6um beads of five different relative intensities, for doing intensity calibrations.
>> Webpage from Thermo Fisher Scientific:
>> https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.t
>> hermofisher.com%2Forder%2Fcatalog%2Fproduct%2FF36909&data=01%7C01%7Cmm
>> odel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018
>> f73e7dd15f26134%7C1&sdata=IbU37oBjbMd9zZE1RKzb4Lp8g6TWlZjE3FJAc6lxtJ0%
>> 3D&reserved=0
>> Manual:
>> https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fasset
>> s.thermofisher.com%2FTFS-Assets%2FLSG%2Fmanuals%2Fmp36909.pdf&data=01%
>> 7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a
>> 1ec44d018f73e7dd15f26134%7C1&sdata=rBOAKsVfx3HvTl0Y2kif8s9lVGzwfgfHYZc
>> %2BYsl1mY0%3D&reserved=0
>>
>> We sell fluorescent beads of smaller sizes, too, if you need them (such as 20 nm) of different wavelengths, for PSF measures.
>>
>> I hope this helps, but please feel free to contact me offline if you wish more info (such as a PowerPoint I have showing images from that last slide).
>>
>> Jason
>>
>>
>> Jason A. Kilgore
>> Technical Application Scientist
>> Molecular Probes / EVOS Tech Support
>> Thermo Fisher Scientific
>>    
>> 1-800-955-6288 then option 4, then option 3, then option 2.
>> Or dial direct at [phone]
>> [hidden email]
>>    
>> This communication is intended solely for the individual/entity to whom it is addressed. It may contain confidential or legally privileged information.  Any unauthorized disclosure or copying is prohibited and may be unlawful. If you have received this communication in error, please notify the sender immediately and delete it from your system.
>>    
>>    
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List
>> [mailto:[hidden email]] On Behalf Of Fco. Javier
>> Díez Guerra
>> Sent: Wednesday, August 30, 2017 5:10 AM
>> To: [hidden email]
>> Subject: Confocal calibration & performance assesment
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.
>> umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cmmodel
>> %40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e
>> 7dd15f26134%7C1&sdata=n6OeQLr2NtHtB320h62kEiK%2BZZJI5PuqSWoqcefDnME%3D
>> &reserved=0 Post images on
>> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cmmodel%40KENT.EDU%7C36cc4a397b8a48289df208d4f22be2af%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=fimjaHIaIMnlqdqyWYm19D0CvjyCBrB08m8x17FX6dk%3D&reserved=0 and include the link in your posting.
>> *****
>>
>> Dear confocalists,
>>
>> I would like to know about the preferred methods used by facility
>> managers to regularly asses the performance of their confocal (and
>> wide-field) microscopes. In other words, which are the most used and reliable methods to check different features of confocal and wide-field microscope peformance?.
>>
>> Particularly, we'd like to check:
>>
>> - illumination uniformity across FOV, now we use a slide coated with fluorescent secondary antibody.
>>
>> - XYZ Chromatic aberration (exciting 405, 488, 561 and 640 nm), now we use 1um beads that can be excited with all laser lines.
>>
>> - XYZ Resolution, we use smaller beads (0,17um) to build and analyze
>> PSFs (for each wavelength)
>>
>> - Laser power and stability: for power, we use a power meter from Newport placed at the objective exit, for stability, we capture long time-series in reflexion mode with an empty preparation.
>>
>> - We do not know how to check detector (PMT, GAsP, Hybrid, etc) sensitivity and SNR.
>>
>> Thanks for your input.
>>
>>
>> --
>> Fco. Javier Diez-Guerra, PhD
>>
>> Profesor Titular UAM
>> Servicio de Microscopía Confocal
>> Centro de Biologia Molecular Severo Ochoa C/ Nicolás Cabrera, 1 Campus
>> de Cantoblanco
>> 28049 Madrid
>> SPAIN
>>
>> Tel     [phone]
>> e-mail: [hidden email]

--


George McNamara, PhD
Baltimore, MD 21231
[hidden email]
https://www.linkedin.com/in/georgemcnamara
https://works.bepress.com/gmcnamara/75   (may need to use Microsoft Edge or Firefox, rather than Google Chrome)
http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650
http://confocal.jhu.edu

July 2017 Current Protocols article, open access:
UNIT 4.4 Microscopy and Image Analysis
http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/abstract
supporting materials direct link is
http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/full#hg0404-sec-0023
figures at
http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/figures

[leavesley]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Another option is Schott filter glass, available from many places. It
happens to be fluorescent and stable.  The different colored filters
provide different fluorescent peak wavelengths.  There was a nice paper
done in Cytometry A on using Schott 475 GG filter glass for routine
benchmarking of fluorescent microscopes:

Halter, Michael, Elianna Bier, Paul C DeRose, Gregory A Cooksey, Steven
J Choquette, Anne L Plant, and John T Elliott. “An Automated Protocol
for Performance Benchmarking a Widefield Fluorescence Microscope.”
/Cytometry Part A/ 85, no. 11 (2014): 978–85.

Best regards,

Silas


On 9/2/2017 12:55 PM, George McNamara wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear listserv,
>
> Fluor-Ref slides ... pick up a box or case from Chroma at a conference.
>
> As for Thermo: how about giving away THIN fluorescent plastic
> specimens .. can include your logo (and no, it would not fall under
> the category of pens or coffee cups that you can no longer give out to
> U.S. academic medical center staff).
>
> Back to Chroma slides ... if not going to 'Neuo' or 'Cell Bio', and
> cannot wait for M&M 8/2018 in Baltimore, as an alternative you can use
> a brightly colored clipboard(s). Come ot think of it, Thermo could
> give out branded clipboards as big reference slides.
>
> enjoy,
>
> George
>
>
> On 9/1/2017 1:26 PM, Kilgore, Jason A. wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your
>> posting.
>> *****
>>
>>
>> ** Vendor response **
>>
>> Hi, Javier,
>>
>> For a uniform fluorescence standard, I recommend the Fluor-Ref
>> slides.  These are sold or re-sold by various places and are fairly
>> inexpensive.  These are plastic slides that are autofluorescent in
>> four different wavelengths and are incredibly uniform and
>> photostable.  The drawbacks are that they are extremely bright, so
>> much so that you often either have to throw in neutral density
>> filters or image an off-wavelength slide. Also, they are thicker than
>> a standard slide.  Here's the webpage from Ted Pella:
>> http://www.tedpella.com/histo_html/fluor.htm.aspx or resold as a set
>> of four from Fisher Scientific:
>> https://www.fishersci.com/shop/products/NC0158348/nc0158348#?keyword=FLUOR-REF+SET+OF+4+SLIDES
>>
>> There are other published methods for uniformity slides (such as a
>> simple solution of fluorescein under a coverslip, or putting dye in
>> agar) or labeling coated slides, but nothing comes close to the
>> Fluor-Ref slides for photostability, storage stability, and
>> uniformity of intensity.
>>
>> There is a good and fairly inexpensive option for some of the other
>> needs you mention:  FocalCheck microspheres slides.  There are
>> different slides, but probably Slide #1 is best for your needs,
>> catalog F36909.  The top row has options where you align rings of
>> different colors to check alignment, or rings and cores of the
>> microspheres. There are also TetraSpeck beads of three different
>> sizes (4, 1, and 0.5 um) that are broadly fluorescent, which you can
>> use for PSF as well as alignment.  The bottom row has 6um beads of
>> five different relative intensities, for doing intensity calibrations.
>> Webpage from Thermo Fisher Scientific:
>> https://www.thermofisher.com/order/catalog/product/F36909
>> Manual:
>> https://assets.thermofisher.com/TFS-Assets/LSG/manuals/mp36909.pdf
>>
>> We sell fluorescent beads of smaller sizes, too, if you need them
>> (such as 20 nm) of different wavelengths, for PSF measures.
>>
>> I hope this helps, but please feel free to contact me offline if you
>> wish more info (such as a PowerPoint I have showing images from that
>> last slide).
>>
>> Jason
>>
>>
>> Jason A. Kilgore
>> Technical Application Scientist
>> Molecular Probes / EVOS Tech Support
>> Thermo Fisher Scientific
>>   1-800-955-6288 then option 4, then option 3, then option 2.
>> Or dial direct at [phone]
>> [hidden email]
>>   This communication is intended solely for the individual/entity to
>> whom it is addressed. It may contain confidential or legally
>> privileged information.  Any unauthorized disclosure or copying is
>> prohibited and may be unlawful. If you have received this
>> communication in error, please notify the sender immediately and
>> delete it from your system.
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List
>> [mailto:[hidden email]] On Behalf Of Fco. Javier
>> Díez Guerra
>> Sent: Wednesday, August 30, 2017 5:10 AM
>> To: [hidden email]
>> Subject: Confocal calibration & performance assesment
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your
>> posting.
>> *****
>>
>> Dear confocalists,
>>
>> I would like to know about the preferred methods used by facility
>> managers to regularly asses the performance of their confocal (and
>> wide-field) microscopes. In other words, which are the most used and
>> reliable methods to check different features of confocal and
>> wide-field microscope peformance?.
>>
>> Particularly, we'd like to check:
>>
>> - illumination uniformity across FOV, now we use a slide coated with
>> fluorescent secondary antibody.
>>
>> - XYZ Chromatic aberration (exciting 405, 488, 561 and 640 nm), now
>> we use 1um beads that can be excited with all laser lines.
>>
>> - XYZ Resolution, we use smaller beads (0,17um) to build and analyze
>> PSFs (for each wavelength)
>>
>> - Laser power and stability: for power, we use a power meter from
>> Newport placed at the objective exit, for stability, we capture long
>> time-series in reflexion mode with an empty preparation.
>>
>> - We do not know how to check detector (PMT, GAsP, Hybrid, etc)
>> sensitivity and SNR.
>>
>> Thanks for your input.
>>
>>
>> --
>> Fco. Javier Diez-Guerra, PhD
>>
>> Profesor Titular UAM
>> Servicio de Microscopía Confocal
>> Centro de Biologia Molecular Severo Ochoa C/ Nicolás Cabrera, 1
>> Campus de Cantoblanco
>> 28049 Madrid
>> SPAIN
>>
>> Tel     [phone]
>> e-mail: [hidden email]
>

--
Silas J. Leavesley, Ph.D.
Professor
Department of Chemical and Biomolecular Engineering
Department of Pharmacology
Center for Lung Biology
University of South Alabama
150 Jaguar Drive, SH4129
Mobile, AL 36688
ph: (251)-460-6160
fax: [phone]
web: http://www.southalabama.edu/centers/bioimaging
google scholar: http://scholar.google.co.uk/citations?user=knkwcj4AAAAJ
------------------------------------------------------------------------

[Jeffrey Carmichael]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

****COMMERCIAL RESPONSE*****

As George says, Chroma Technology always gives these away at meetings.

However, we also sell them
<https://www.chroma.com/products/diagnostic-slides> (catalog #92001).
They're the same slides as the aforementioned for sale through other
vendors.  $20.  Thinner versions may be coming......

Jeff


*Jeff Carmichael*

*Technical and Product Marketing Manager**[hidden email]
<[hidden email]>*

On Sat, Sep 2, 2017 at 1:55 PM, George McNamara <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear listserv,
>
> Fluor-Ref slides ... pick up a box or case from Chroma at a conference.
>
> As for Thermo: how about giving away THIN fluorescent plastic specimens ..
> can include your logo (and no, it would not fall under the category of pens
> or coffee cups that you can no longer give out to U.S. academic medical
> center staff).
>
> Back to Chroma slides ... if not going to 'Neuo' or 'Cell Bio', and cannot
> wait for M&M 8/2018 in Baltimore, as an alternative you can use a brightly
> colored clipboard(s). Come ot think of it, Thermo could give out branded
> clipboards as big reference slides.
>
> enjoy,
>
> George
>
>
> On 9/1/2017 1:26 PM, Kilgore, Jason A. wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>>
>> ** Vendor response **
>>
>> Hi, Javier,
>>
>> For a uniform fluorescence standard, I recommend the Fluor-Ref slides.
>> These are sold or re-sold by various places and are fairly inexpensive.
>> These are plastic slides that are autofluorescent in four different
>> wavelengths and are incredibly uniform and photostable.  The drawbacks are
>> that they are extremely bright, so much so that you often either have to
>> throw in neutral density filters or image an off-wavelength slide.  Also,
>> they are thicker than a standard slide.  Here's the webpage from Ted
>> Pella:  http://www.tedpella.com/histo_html/fluor.htm.aspx or resold as a
>> set of four from Fisher Scientific: https://www.fishersci.com/shop
>> /products/NC0158348/nc0158348#?keyword=FLUOR-REF+SET+OF+4+SLIDES
>>
>> There are other published methods for uniformity slides (such as a simple
>> solution of fluorescein under a coverslip, or putting dye in agar) or
>> labeling coated slides, but nothing comes close to the Fluor-Ref slides for
>> photostability, storage stability, and uniformity of intensity.
>>
>> There is a good and fairly inexpensive option for some of the other needs
>> you mention:  FocalCheck microspheres slides.  There are different slides,
>> but probably Slide #1 is best for your needs, catalog F36909.  The top row
>> has options where you align rings of different colors to check alignment,
>> or rings and cores of the microspheres. There are also TetraSpeck beads of
>> three different sizes (4, 1, and 0.5 um) that are broadly fluorescent,
>> which you can use for PSF as well as alignment.  The bottom row has 6um
>> beads of five different relative intensities, for doing intensity
>> calibrations.
>> Webpage from Thermo Fisher Scientific:  https://www.thermofisher.com/o
>> rder/catalog/product/F36909
>> Manual: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/mp369
>> 09.pdf
>>
>> We sell fluorescent beads of smaller sizes, too, if you need them (such
>> as 20 nm) of different wavelengths, for PSF measures.
>>
>> I hope this helps, but please feel free to contact me offline if you wish
>> more info (such as a PowerPoint I have showing images from that last slide).
>>
>> Jason
>>
>>
>> Jason A. Kilgore
>> Technical Application Scientist
>> Molecular Probes / EVOS Tech Support
>> Thermo Fisher Scientific
>>   1-800-955-6288 then option 4, then option 3, then option 2.
>> Or dial direct at [phone]
>> [hidden email]
>>   This communication is intended solely for the individual/entity to whom
>> it is addressed. It may contain confidential or legally privileged
>> information.  Any unauthorized disclosure or copying is prohibited and may
>> be unlawful. If you have received this communication in error, please
>> notify the sender immediately and delete it from your system.
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List [mailto:[hidden email]]
>> On Behalf Of Fco. Javier Díez Guerra
>> Sent: Wednesday, August 30, 2017 5:10 AM
>> To: [hidden email]
>> Subject: Confocal calibration & performance assesment
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Dear confocalists,
>>
>> I would like to know about the preferred methods used by facility
>> managers to regularly asses the performance of their confocal (and
>> wide-field) microscopes. In other words, which are the most used and
>> reliable methods to check different features of confocal and wide-field
>> microscope peformance?.
>>
>> Particularly, we'd like to check:
>>
>> - illumination uniformity across FOV, now we use a slide coated with
>> fluorescent secondary antibody.
>>
>> - XYZ Chromatic aberration (exciting 405, 488, 561 and 640 nm), now we
>> use 1um beads that can be excited with all laser lines.
>>
>> - XYZ Resolution, we use smaller beads (0,17um) to build and analyze PSFs
>> (for each wavelength)
>>
>> - Laser power and stability: for power, we use a power meter from Newport
>> placed at the objective exit, for stability, we capture long time-series in
>> reflexion mode with an empty preparation.
>>
>> - We do not know how to check detector (PMT, GAsP, Hybrid, etc)
>> sensitivity and SNR.
>>
>> Thanks for your input.
>>
>>
>> --
>> Fco. Javier Diez-Guerra, PhD
>>
>> Profesor Titular UAM
>> Servicio de Microscopía Confocal
>> Centro de Biologia Molecular Severo Ochoa C/ Nicolás Cabrera, 1 Campus de
>> Cantoblanco
>> 28049 Madrid
>> SPAIN
>>
>> Tel     [phone]
>> e-mail: [hidden email]
>>
>
> --
>
>
> George McNamara, PhD
> Baltimore, MD 21231
> [hidden email]
> https://www.linkedin.com/in/georgemcnamara
> https://works.bepress.com/gmcnamara/75   (may need to use Microsoft Edge
> or Firefox, rather than Google Chrome)
> http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650
> http://confocal.jhu.edu
>
> July 2017 Current Protocols article, open access:
> UNIT 4.4 Microscopy and Image Analysis
> http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/abstract
> supporting materials direct link is
> http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/full#hg0404-sec-0023
> figures at
> http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/figures
>

--

<https://www.chroma.com/> CHROMA TECHNOLOGY CORP®
*an employee owned company*
10 Imtec Lane, Bellows Falls, Vermont 05101 USA
800-824-7662 | 802-428-2525
www.chroma.com | [hidden email]                            

[Barbara Foster]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Listers,

Actually, Fluor-Ref slides were invented by Dr.
P. C. Cheng (SUNY/Buffalo), originally for use in
his classroom.  Because of our activities in
microscopy education, P. C. asked us to carry
them . and, to the best of my knowledge,
Microscopy/Microscopy Education
(MicroscopyEducation.com) is the only recognized commercial source.

We agree with Jason, they are bright, but that
problem is readily solved using ND filters.  To
retain consistency from batch to batch, we've
opted to continue with his original formulation
for the original set of 4 colors (blue, green,
yellow, and red excitation).  However, if you are
interested, we do have a limited supply of  "10%
intensity" slides that excite similarly to FITC.

And Geoge is correct... they are inexpensive when
compared to other reference slides.  They are
plastic, but they are durable and have been used
by thousands of facilities world wide

Spectra, FAQs, etc. are on our website, www.microscopyeducation.com.

***CAVEAT** Commercial posting

Good Hunting!
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education  ... "Education, not JustTraining"
7101 Royal Glen Trail, Suite A  - McKinney, TX 75070
P: 972-924-5310 W: www.MicroscopyEducation.com
(Microscopy/Microscopy Education is a division of
The Microscopy & Imaging Place, Inc.)

NEW!   Getting involved in Raman or FTIR?
MME is now offering courses in these areas
specifically for microscopists!  We can customize
a course on nearly any topic, from fluorescence
to confocal to image analysis to SEM/TEM.  Call today for details.









At 01:52 PM 9/2/2017, George McNamara wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com and include the link in your posting.
>*****
>
>Dear listserv,
>
>Fluor-Ref slides ... pick up a box or case from Chroma at a conference.
>
>As for Thermo: how about giving away THIN
>fluorescent plastic specimens .. can include
>your logo (and no, it would not fall under the
>category of pens or coffee cups that you can no
>longer give out to U.S. academic medical center staff).
>
>Back to Chroma slides ... if not going to 'Neuo'
>or 'Cell Bio', and cannot wait for M&M 8/2018 in
>Baltimore, as an alternative you can use a
>brightly colored clipboard(s). Come ot think of
>it, Thermo could give out branded clipboards as big reference slides.
>
>enjoy,
>
>George
>
>
>On 9/1/2017 1:26 PM, Kilgore, Jason A. wrote:
>>*****
>>To join, leave or search the confocal microscopy listserv, go to:
>>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>Post images on http://www.imgur.com and include the link in your posting.
>>*****
>>
>>
>>** Vendor response **
>>
>>Hi, Javier,
>>
>>For a uniform fluorescence standard, I
>>recommend the Fluor-Ref slides.  These are sold
>>or re-sold by various places and are fairly
>>inexpensive.  These are plastic slides that are
>>autofluorescent in four different wavelengths
>>and are incredibly uniform and
>>photostable.  The drawbacks are that they are
>>extremely bright, so much so that you often
>>either have to throw in neutral density filters
>>or image an off-wavelength slide.  Also, they
>>are thicker than a standard slide.  Here's the
>>webpage from Ted
>>Pella:
>>http://www.tedpella.com/histo_html/fluor.htm.asp 
>>x or resold as a set of four from Fisher
>>Scientific:
>>https://www.fishersci.com/shop/products/NC0158348/nc0158348#?keyword=FLUOR-REF+SET+OF+4+SLIDES
>>
>>There are other published methods for
>>uniformity slides (such as a simple solution of
>>fluorescein under a coverslip, or putting dye
>>in agar) or labeling coated slides, but nothing
>>comes close to the Fluor-Ref slides for
>>photostability, storage stability, and uniformity of intensity.
>>
>>There is a good and fairly inexpensive option
>>for some of the other needs you
>>mention:  FocalCheck microspheres
>>slides.  There are different slides, but
>>probably Slide #1 is best for your needs,
>>catalog F36909.  The top row has options where
>>you align rings of different colors to check
>>alignment, or rings and cores of the
>>microspheres. There are also TetraSpeck beads
>>of three different sizes (4, 1, and 0.5 um)
>>that are broadly fluorescent, which you can use
>>for PSF as well as alignment.  The bottom row
>>has 6um beads of five different relative
>>intensities, for doing intensity calibrations.
>>Webpage from Thermo Fisher
>>Scientific:  https://www.thermofisher.com/order/catalog/product/F36909
>>Manual: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/mp36909.pdf
>>
>>We sell fluorescent beads of smaller sizes,
>>too, if you need them (such as 20 nm) of
>>different wavelengths, for PSF measures.
>>
>>I hope this helps, but please feel free to
>>contact me offline if you wish more info (such
>>as a PowerPoint I have showing images from that last slide).
>>
>>Jason
>>
>>
>>Jason A. Kilgore
>>Technical Application Scientist
>>Molecular Probes / EVOS Tech Support
>>Thermo Fisher Scientific
>>
>>1-800-955-6288 then option 4, then option 3, then option 2.
>>Or dial direct at [phone]
>>[hidden email]
>>
>>This communication is intended solely for the
>>individual/entity to whom it is addressed. It
>>may contain confidential or legally privileged
>>information.  Any unauthorized disclosure or
>>copying is prohibited and may be unlawful. If
>>you have received this communication in error,
>>please notify the sender immediately and delete it from your system.
>>
>>
>>
>>
>>-----Original Message-----
>>From: Confocal Microscopy List
>>[mailto:[hidden email]] On Behalf Of Fco. Javier Díez Guerra
>>Sent: Wednesday, August 30, 2017 5:10 AM
>>To: [hidden email]
>>Subject: Confocal calibration & performance assesment
>>
>>*****
>>To join, leave or search the confocal microscopy listserv, go to:
>>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>Post images on http://www.imgur.com and include the link in your posting.
>>*****
>>
>>Dear confocalists,
>>
>>I would like to know about the preferred
>>methods used by facility managers to regularly
>>asses the performance of their confocal (and
>>wide-field) microscopes. In other words, which
>>are the most used and reliable methods to check
>>different features of confocal and wide-field microscope peformance?.
>>
>>Particularly, we'd like to check:
>>
>>- illumination uniformity across FOV, now we
>>use a slide coated with fluorescent secondary antibody.
>>
>>- XYZ Chromatic aberration (exciting 405, 488,
>>561 and 640 nm), now we use 1um beads that can be excited with all laser lines.
>>
>>- XYZ Resolution, we use smaller beads (0,17um)
>>to build and analyze PSFs (for each wavelength)
>>
>>- Laser power and stability: for power, we use
>>a power meter from Newport placed at the
>>objective exit, for stability, we capture long
>>time-series in reflexion mode with an empty preparation.
>>
>>- We do not know how to check detector (PMT,
>>GAsP, Hybrid, etc) sensitivity and SNR.
>>
>>Thanks for your input.
>>
>>
>>--
>>Fco. Javier Diez-Guerra, PhD
>>
>>Profesor Titular UAM
>>Servicio de Microscopía Confocal
>>Centro de Biologia Molecular Severo Ochoa C/
>>Nicolás Cabrera, 1 Campus de Cantoblanco
>>28049 Madrid
>>SPAIN
>>
>>Tel     [phone]
>>e-mail: [hidden email]
>
>--
>
>
>George McNamara, PhD
>Baltimore, MD 21231
>[hidden email]
>https://www.linkedin.com/in/georgemcnamara
>https://works.bepress.com/gmcnamara/75  (may
>need to use Microsoft Edge or Firefox, rather than Google Chrome)
>http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650
>http://confocal.jhu.edu
>
>July 2017 Current Protocols article, open access:
>UNIT 4.4 Microscopy and Image Analysis
>http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/abstract
>supporting materials direct link is
>http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/full#hg0404-sec-0023
>figures at
>http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/figures