Structured Illumination vs Deconvolution
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Claire Brown |
Structured Illumination vs Deconvolution
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire |
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Patrick Van Oostveldt |
Re: Structured Illumination vs Deconvolution
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Yes it can if we refer to Lothar Schermelleh et al. Science 320, p1332 (2008) and Gustafsson et al. Biophysical J. 94, p4957 (2008) The only bottleneck is time and datacongestion. (2 min for a 512x512x80 volume and 2 min calculation with this) . So live imaging is nearly impossible I guess, but it should be nice if the authors could comment in this. Patrick Quoting Claire Brown <[hidden email]>: > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > I was wondering if there is any clear advantage to structured illumination > versus deconvolution? Can it actually give you higher resolution? > > Sincerely, > > Claire > -- Dep. Moleculaire Biotechnologie Coupure links 653 B 9000 GENT tel 09 264 5969 fax 09 264 6219 |
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Steffen Steinert |
AW: Structured Illumination vs Deconvolution
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In reply to this post by Claire Brown
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Claire, According to recent reports (Schermelleh et al., Science 2008 or Gustafsson et al., Biophysical Journal 2008) Widefield-SIM breaks the resolution by a factor of about 2 to 3, laterally AND axially. Thus, you can separate objects being apart 100nm in lateral direction and 300nm axially. From my understanding, the restoration of attenuated higher spatial frequencies due to Deconvolution of 'ordinary' Widefield images will give you at best a resolution proposed by Abbe/Rayleigh (in frequency domain k(z)= NA²/2ηλ and k(xy)=2NA/λ). As far as I know (and I know rather little), calculating information beyond that limit using Deconvolution is not possible, since your OTF is zero and hence the information is lost and not just attenuated. Due to the spatial frequency mixing with a certain illumination pattern in SIM (interference of 3 mutually coherent beams in the Gustafsson paper) you will get a way bigger total Optical-Transfer-Function (with higher k-values) since you can additively overlay these OTFs in the frequency domain. Therefore, you´ll obtain images with a higher resolution than with a Widefield/Deconvolution system. Hope this helps, Cheers, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Claire Brown Gesendet: Freitag, 27. Juni 2008 16:51 An: [hidden email] Betreff: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire |
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Steffen Steinert |
AW: Structured Illumination vs Deconvolution
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In reply to this post by Claire Brown
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Claire, According to recent reports (e.g. Schermelleh, Science 2008 or Gustafsson, Biophysical Journal 2008) Widefield-SIM breaks the resolution by a factor of about 2 to 3, laterally AND axially. Thus, you can separate objects being apart 100nm in lateral direction and 300nm axially. From my understanding, the restoration of attenuated higher spatial frequencies due to Deconvolution of 'ordinary' Widefield images will give you at best a resolution proposed by Abbe/Rayleigh (in frequency domain k(z)= NA²/2ηλ and k(xy)=2NA/λ). As far as I know (and I know rather little), calculating information beyond that limit using Deconvolution is not possible, since your OTF is zero and hence the information is lost and not just attenuated. Due to the spatial frequency mixing with a certain illumination pattern in SIM (interference of 3 mutually coherent beams in the Gustafsson paper) you will get a way bigger total Optical-Transfer-Function (with higher k-values) since you can additively overlay these OTFs in the frequency domain. Therefore, you´ll obtain images with a higher resolution than with a Widefield/Deconvolution system. Hope this helps, Cheers, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire |
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Steffen Steinert |
AW: Structured Illumination vs Deconvolution
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In reply to this post by Claire Brown
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Claire, According to recent reports Widefield-SIM breaks the resolution by a factor of about 2 to 3, laterally AND axially. Thus, you can separate objects being apart 100nm in lateral direction and 300nm axially. From my understanding, the restoration of attenuated higher spatial frequencies due to Deconvolution of 'ordinary' Widefield images will give you at best a resolution proposed by Abbe/Rayleigh (in frequency domain k(z)= NA²/2ηλ and k(xy)=2NA/λ). As far as I know (and I know rather little), calculating information beyond that limit using Deconvolution is not possible, since your OTF is zero and hence the information is lost and not just attenuated. Due to the spatial frequency mixing with a certain illumination pattern in SIM (interference of 3 mutually coherent beams in the Gustafsson paper Biophysical Journal 2008 or Schermelleh, Science 2008) you will get a way bigger total Optical-Transfer-Function (with higher k-values) since you can additively overlay these OTFs in the frequency domain. Therefore, you´ll obtain images with a higher resolution than with a Widefield/Deconvolution system. Hope this helps, Cheers, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Claire Brown Gesendet: Freitag, 27. Juni 2008 16:51 An: [hidden email] Betreff: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire |
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Steffen Steinert |
AW: Structured Illumination vs Deconvolution
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In reply to this post by Claire Brown
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Claire, *The list always rejects my email, perhaps this one gets through... * According to recent reports Widefield-SIM (e.g. Schermelleh, Science 2008 or Gustafsson, Biophysical Journal 2008) breaks the resolution by a factor of about 2 to 3, laterally AND axially. Thus, you can separate objects being apart 100nm in lateral direction and 300nm axially. From my understanding, the restoration of attenuated higher spatial frequencies due to Deconvolution of 'ordinary' Widefield images will give you at best a resolution proposed by Abbe/Rayleigh (in frequency domain k(z)= NA²/2ηλ and k(xy)=2NA/λ). As far as I know (and I know rather little), calculating information beyond that limit using Deconvolution is not possible, since your OTF is zero and hence the information is lost and not just attenuated. Due to the spatial frequency mixing with a certain illumination pattern in SIM (interference of 3 mutually coherent beams in the Gustafsson paper) you will get a way bigger total Optical-Transfer-Function (with higher k-values) since you can additively overlay these OTFs in the frequency domain. Therefore, in terms of resolution you´ll obtain better images with SIM than with a Widefield/Deconvolution system. Hope this helps, Cheers, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Claire Brown Gesendet: Freitag, 27. Juni 2008 16:51 An: [hidden email] Betreff: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire |
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Steffen Steinert |
AW: Structured Illumination vs Deconvolution
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In reply to this post by Claire Brown
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Claire, *The list always rejects my email, perhaps this one gets through... * According to recent reports Widefield-SIM breaks the resolution by a factor of about 2 to 3, laterally AND axially. Thus, you can separate objects being apart 100nm in lateral direction and 300nm axially. From my understanding, the restoration of attenuated higher spatial frequencies due to Deconvolution of 'ordinary' Widefield images will give you at best a resolution proposed by Abbe/Rayleigh (in frequency domain k(z)= NA²/2ηλ and k(xy)=2NA/λ). As far as I know (and I know rather little), calculating information beyond that limit using Deconvolution is not possible, since your OTF is zero and hence the information is lost and not just attenuated. Due to the spatial frequency mixing with a certain illumination pattern in SIM (interference of 3 mutually coherent beams in the Gustafsson or Schermelleh paper) you will get a way bigger total Optical-Transfer-Function (with higher k-values) since you can additively overlay these OTFs in the frequency domain. Therefore, in terms of resolution you´ll obtain better images with SIM than with a Widefield/Deconvolution system. Hope this helps, Cheers, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Claire Brown Gesendet: Freitag, 27. Juni 2008 16:51 An: [hidden email] Betreff: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire |
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Steffen Steinert |
AW: Structured Illumination vs Deconvolution
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In reply to this post by Steffen Steinert
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Sorry, for the multiple sending of the email. I always got an response from the list, that my email has been rejected so I sent it again without knowing it´s been sent already. SORRY, won´t happen again! Steffen -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Steffen Steinert Gesendet: Freitag, 27. Juni 2008 18:30 An: [hidden email] Betreff: AW: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Claire, According to recent reports (Schermelleh et al., Science 2008 or Gustafsson et al., Biophysical Journal 2008) Widefield-SIM breaks the resolution by a factor of about 2 to 3, laterally AND axially. Thus, you can separate objects being apart 100nm in lateral direction and 300nm axially. >From my understanding, the restoration of attenuated higher spatial frequencies due to Deconvolution of 'ordinary' Widefield images will give you at best a resolution proposed by Abbe/Rayleigh (in frequency domain k(z)= NA²/2ηλ and k(xy)=2NA/λ). As far as I know (and I know rather little), calculating information beyond that limit using Deconvolution is not possible, since your OTF is zero and hence the information is lost and not just attenuated. Due to the spatial frequency mixing with a certain illumination pattern in SIM (interference of 3 mutually coherent beams in the Gustafsson paper) you will get a way bigger total Optical-Transfer-Function (with higher k-values) since you can additively overlay these OTFs in the frequency domain. Therefore, you´ll obtain images with a higher resolution than with a Widefield/Deconvolution system. Hope this helps, Cheers, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Claire Brown Gesendet: Freitag, 27. Juni 2008 16:51 An: [hidden email] Betreff: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire |
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Ignatius, Mike |
Rejected postings warnings keep coming up.
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I have been getting similar odd notification each time I submit of late. Wondering if a fix is needed.
Starts of with subject line: Rejected posting to [hidden email] Then this subject: "Your message is being returned to you unprocessed because it appears to have already been distributed to the CONFOCAL list. That is, a message with identical text (but possibly with different mail headers) has been posted to the list recently, either by you or by someone else. If you have reason to resend this message to the list (for instance because you have been notified of a hardware failure with loss of data), please alter the text of the message in some way and resend it to the list. Altering the "Subject:" line or adding blank lines at the top or bottom of the message is not sufficient. Instead, you should add a sentence or two at the top explaining why you are resending the message. This explanation will help the other subscribers understand why they are getting two copies of the same message." I haven't been double posting either. Ideas on how to remedy? Mike Ignatius, Molecular Probes/Invitrogen -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Steffen Steinert Sent: Friday, June 27, 2008 9:57 AM To: [hidden email] Subject: AW: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Sorry, for the multiple sending of the email. I always got an response from the list, that my email has been rejected so I sent it again without knowing it´s been sent already. SORRY, won´t happen again! Steffen -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Steffen Steinert Gesendet: Freitag, 27. Juni 2008 18:30 An: [hidden email] Betreff: AW: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Claire, According to recent reports (Schermelleh et al., Science 2008 or Gustafsson et al., Biophysical Journal 2008) Widefield-SIM breaks the resolution by a factor of about 2 to 3, laterally AND axially. Thus, you can separate objects being apart 100nm in lateral direction and 300nm axially. >From my understanding, the restoration of attenuated higher spatial frequencies due to Deconvolution of 'ordinary' Widefield images will give you at best a resolution proposed by Abbe/Rayleigh (in frequency domain k(z)= NA²/2ηλ and k(xy)=2NA/λ). As far as I know (and I know rather little), calculating information beyond that limit using Deconvolution is not possible, since your OTF is zero and hence the information is lost and not just attenuated. Due to the spatial frequency mixing with a certain illumination pattern in SIM (interference of 3 mutually coherent beams in the Gustafsson paper) you will get a way bigger total Optical-Transfer-Function (with higher k-values) since you can additively overlay these OTFs in the frequency domain. Therefore, you´ll obtain images with a higher resolution than with a Widefield/Deconvolution system. Hope this helps, Cheers, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Claire Brown Gesendet: Freitag, 27. Juni 2008 16:51 An: [hidden email] Betreff: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire |
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Barbara Foster |
Re: AW: Structured Illumination vs Deconvolution - Another Alternative
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In reply to this post by Steffen Steinert
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear Listers,
There is a third alternative. I just got back yesterday from a private meeting hosted by CytoViva. They have been developing a series of very interesting applications on live cell imaging, both as a single imaging modality and combined with fluorescence. Although they call their device a "darkfield condenser," the behavior I have seen far exceed darkfield microscopy in several ways: (a) while darkfield provides unlimited depth of field, CytoViva provides elegant, discrete optical sectioning (b) while darkfield is detection limited, tests with a Richardson resolution target indicate that CytoViva produces true resolution on the order of 90-92 nm. (c) while darkfield improves the S/N ratio, CytoViva dramatically improves S/N. This enhanced S/N, combined with its higher resolution makes it an great solution for live imaging of very tiny point-like objects such as nano gold particles. To date, I have not seen a satisfactory scientific explanation as to why their system works this way... it just really works. (For the gurus on the listserver: If you want to have a discussion, please email me or give me a call offline (972-924-5310). I've got some ideas about what's going on, but would benefit from your broader experience). Anyway, when you put all of these attributes together, you get high signal/high resolution (beyond the Abbe limit) imaging of live cells without staining. When you add fluorescence (with their "DFM module"), you can produce a wonderful, live cell image of the cellular context simultaneously with the chemical tagged fluorescence ... all real-time, without the need to overlay computer images. So, for those of you who need to know exactly where a tagged molecule/drug, etc. is going in a cell, and, especially for those of you who want to document the process over time, this system provides an excellent solution at a reasonable price tag. (I think that the combined CytoViva/Fluorescence module combo ... including all the necessary light sources, etc., is on the order of 25K. They now have both upright and inverted versions that fit most microscopes). At the meeting, they also discussed their new Hyperspectral Imaging (HSI) system. While their condenser/illumination provides excellent high resolution/high S to N qualitative images, the HSI is able to collect a visible/near IR spectrum to show quantitative chemical shifts, either over the whole image (pixel by pixel) or in selected ROIs. This product is being finalized and will be formally announced later this year. In the interim, the folks at CytoViva are actively developing their application expertise, so if you think this would be of value in your lab, I encourage you to contact the head of Technical Sales: Byron Cheatham ([hidden email]). For those of you who are just curious, I encourage you to visit their website: www.CytoViva.com . They have some great videos, both of the instrumentation and applications. Caveat: while I launched the original CytoViva in 2004, I currently have no financial interest in this product. It's just neat technology. Hope this is helpful! Best regards, Barbara Foster, President Microscopy/Microscopy Education 7101 Royal Glen Trail, Suite A ~ McKinney TX 75070 ~ P: (972)924-5310 ~ Skype: fostermme ~ W: www.MicroscopyEducation.com NEWS! Our website, which had been down for nearly a month due to hackers, is back and better than ever. Come visit us at www.MicroscopyEducation.com! And Don't forget: MME is now scheduling customized, on-site courses through Dec 2008. Call me for details. At 04:36 AM 6/27/2008, Steffen Steinert wrote: Search the CONFOCAL archive at |
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Loralei Dewe-2 |
Re: Structured Illumination vs Deconvolution
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In reply to this post by Steffen Steinert
Does anyone have any of these images they would be willing to share? I am very interested in seeing how this works?
If you would be willing to share please contact me privately ;-) Cheers, Loralei Dewe Applications Specialist 5916 Sunnybrook Lane Dixon, Ca 95620 Phone: 707-446-8759 Fax: 707-446-8759 *51 Cell: 707-301-0604 MAG Biosystems | Photometrics | QImaging | Media Cybernetics | Gatan www.MAGbiosystems.com www.photomet.com www.qimaging.com www.mediacy.com www.gatan.com Microimaging Applications Group (MAG) The Microimaging Applications Group comprises five imaging leaders: Photometrics, Gatan, Media Cybernetics, QImaging, and MAG Biosystems. These technology partners work independently as well as in synergy to offer an unparalleled range of solutions for microimaging applications. This message and any attachments are solely for the use of intended recipients. They may contain privileged and/or confidential information. If you are not the intended recipient, you are hereby notified that you received this email in error, and that any review, dissemination, distribution or copying of this email and any attachment is strictly prohibited. If you receive this email in error please contact the sender and delete the message and any attachments associated therewith from your computer. Your cooperation in this matter is appreciated. -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Steffen Steinert Sent: Friday, June 27, 2008 8:57 AM To: [hidden email] Subject: AW: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Sorry, for the multiple sending of the email. I always got an response from the list, that my email has been rejected so I sent it again without knowing it´s been sent already. SORRY, won´t happen again! Steffen -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Steffen Steinert Gesendet: Freitag, 27. Juni 2008 18:30 An: [hidden email] Betreff: AW: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Claire, According to recent reports (Schermelleh et al., Science 2008 or Gustafsson et al., Biophysical Journal 2008) Widefield-SIM breaks the resolution by a factor of about 2 to 3, laterally AND axially. Thus, you can separate objects being apart 100nm in lateral direction and 300nm axially. >From my understanding, the restoration of attenuated higher spatial frequencies due to Deconvolution of 'ordinary' Widefield images will give you at best a resolution proposed by Abbe/Rayleigh (in frequency domain k(z)= NA²/2ηλ and k(xy)=2NA/λ). As far as I know (and I know rather little), calculating information beyond that limit using Deconvolution is not possible, since your OTF is zero and hence the information is lost and not just attenuated. Due to the spatial frequency mixing with a certain illumination pattern in SIM (interference of 3 mutually coherent beams in the Gustafsson paper) you will get a way bigger total Optical-Transfer-Function (with higher k-values) since you can additively overlay these OTFs in the frequency domain. Therefore, you´ll obtain images with a higher resolution than with a Widefield/Deconvolution system. Hope this helps, Cheers, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Claire Brown Gesendet: Freitag, 27. Juni 2008 16:51 An: [hidden email] Betreff: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire |
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Klughammer Industrie GmbH |
Re: Structured Illumination vs Deconvolution
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In reply to this post by Claire Brown
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Claire, it depends whether you want to know more about 1) three-dimensional structured illumination microscopy (3D-SIM) see: http://www.cipsm.de/en/publications/researchAreaD/Subdiffraction_Multicolor_Imaging/index.html?style=0 or 2) structured illumination system like ApoTome or OptiGrid see: http://listserv.buffalo.edu/cgi-bin/wa?A3=ind0403&L=CONFOCAL&E=0&P=117010&B=--%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D_-1133271579%3D%3D_ma%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D&T=text%2Fhtml;%20charset=us-ascii Kind regards Anneliese 2008/6/27 Claire Brown <[hidden email]>: > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > I was wondering if there is any clear advantage to structured illumination > versus deconvolution? Can it actually give you higher resolution? > > Sincerely, > > Claire > |
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Structured illumination can do all sorts of things - it just depends what your goal is. Simple structured illumination systems such as Apotome or Optigrid will give you optical sectioning. 'Clever' sysyems - mostly based on the work of Mats Gustafsson - can give you a two-fold resolution improvement which is not to be sneezed at. When you get into saturated pattern structured illumination the resolution (as in STED) is in principle unlimited (in practice, I guess, around 90 nm). Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Klughammer Industrie GmbH Sent: Saturday, 28 June 2008 10:45 PM To: [hidden email] Subject: Re: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Claire, it depends whether you want to know more about 1) three-dimensional structured illumination microscopy (3D-SIM) see: http://www.cipsm.de/en/publications/researchAreaD/Subdiffraction_Multicolor_Imaging/index.html?style=0 or 2) structured illumination system like ApoTome or OptiGrid see: http://listserv.buffalo.edu/cgi-bin/wa?A3=ind0403&L=CONFOCAL&E=0&P=117010&B=--%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D_-1133271579%3D%3D_ma%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D&T=text%2Fhtml;%20charset=us-ascii Kind regards Anneliese 2008/6/27 Claire Brown <[hidden email]>: > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > I was wondering if there is any clear advantage to structured > illumination versus deconvolution? Can it actually give you higher resolution? > > Sincerely, > > Claire > No virus found in this incoming message. Checked by AVG. Version: 7.5.526 / Virus Database: 270.4.1/1522 - Release Date: 27/06/2008 8:27 AM No virus found in this outgoing message. Checked by AVG. Version: 7.5.526 / Virus Database: 270.4.1/1522 - Release Date: 27/06/2008 8:27 AM |
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In reply to this post by Ignatius, Mike
The problem appears to be that for some reason every message
gets submitted in duplicate. This naturally causes the server to reject the second copy on the grounds it has already been posted (which it has). What needs to be sorted is why messages are getting posted twice when they are only sent once. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ignatius, Mike Sent: Saturday, 28 June 2008 3:12 AM To: [hidden email] Subject: Rejected postings warnings keep coming up. I have been getting similar odd notification each time I submit of late. Wondering if a fix is needed. Starts of with subject line: Rejected posting to [hidden email] Then this subject: "Your message is being returned to you unprocessed because it appears to have already been distributed to the CONFOCAL list. That is, a message with identical text (but possibly with different mail headers) has been posted to the list recently, either by you or by someone else. If you have reason to resend this message to the list (for instance because you have been notified of a hardware failure with loss of data), please alter the text of the message in some way and resend it to the list. Altering the "Subject:" line or adding blank lines at the top or bottom of the message is not sufficient. Instead, you should add a sentence or two at the top explaining why you are resending the message. This explanation will help the other subscribers understand why they are getting two copies of the same message." I haven't been double posting either. Ideas on how to remedy? Mike Ignatius, Molecular Probes/Invitrogen -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Steffen Steinert Sent: Friday, June 27, 2008 9:57 AM To: [hidden email] Subject: AW: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Sorry, for the multiple sending of the email. I always got an response from the list, that my email has been rejected so I sent it again without knowing it´s been sent already. SORRY, won´t happen again! Steffen -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Steffen Steinert Gesendet: Freitag, 27. Juni 2008 18:30 An: [hidden email] Betreff: AW: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Claire, According to recent reports (Schermelleh et al., Science 2008 or Gustafsson et al., Biophysical Journal 2008) Widefield-SIM breaks the resolution by a factor of about 2 to 3, laterally AND axially. Thus, you can separate objects being apart 100nm in lateral direction and 300nm axially. >From my understanding, the restoration of attenuated higher spatial frequencies due to Deconvolution of 'ordinary' Widefield images will give you at best a resolution proposed by Abbe/Rayleigh (in frequency domain k(z)= NA²/2ηλ and k(xy)=2NA/λ). As far as I know (and I know rather little), calculating information beyond that limit using Deconvolution is not possible, since your OTF is zero and hence the information is lost and not just attenuated. Due to the spatial frequency mixing with a certain illumination pattern in SIM (interference of 3 mutually coherent beams in the Gustafsson paper) you will get a way bigger total Optical-Transfer-Function (with higher k-values) since you can additively overlay these OTFs in the frequency domain. Therefore, you´ll obtain images with a higher resolution than with a Widefield/Deconvolution system. Hope this helps, Cheers, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- -----Ursprüngliche Nachricht----- Von: Confocal Microscopy List [mailto:[hidden email]] Im Auftrag von Claire Brown Gesendet: Freitag, 27. Juni 2008 16:51 An: [hidden email] Betreff: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I was wondering if there is any clear advantage to structured illumination versus deconvolution? Can it actually give you higher resolution? Sincerely, Claire No virus found in this incoming message. Checked by AVG. Version: 7.5.524 / Virus Database: 270.4.1/1519 - Release Date: 25/06/2008 4:13 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.526 / Virus Database: 270.4.1/1522 - Release Date: 27/06/2008 8:27 AM |
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Alberto Diaspro |
R: Re: Structured Illumination vs Deconvolution
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In reply to this post by Guy Cox
You can get 20 30 With excellent 'steadable' molecules i guess alby
Le mail ti raggiungono ovunque con BlackBerry® from Vodafone! -----Original Message----- From: Guy Cox <[hidden email]> Date: Sun, 29 Jun 2008 23:32:26 To: <[hidden email]> Subject: Re: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Structured illumination can do all sorts of things - it just depends what your goal is. Simple structured illumination systems such as Apotome or Optigrid will give you optical sectioning. 'Clever' sysyems - mostly based on the work of Mats Gustafsson - can give you a two-fold resolution improvement which is not to be sneezed at. When you get into saturated pattern structured illumination the resolution (as in STED) is in principle unlimited (in practice, I guess, around 90 nm). Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Klughammer Industrie GmbH Sent: Saturday, 28 June 2008 10:45 PM To: [hidden email] Subject: Re: Structured Illumination vs Deconvolution Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Claire, it depends whether you want to know more about 1) three-dimensional structured illumination microscopy (3D-SIM) see: http://www.cipsm.de/en/publications/researchAreaD/Subdiffraction_Multicolor_Imaging/index.html?style=0 or 2) structured illumination system like ApoTome or OptiGrid see: http://listserv.buffalo.edu/cgi-bin/wa?A3=ind0403&L=CONFOCAL&E=0&P=117010&B=--%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D_-1133271579%3D%3D_ma%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D%3D&T=text%2Fhtml;%20charset=us-ascii Kind regards Anneliese 2008/6/27 Claire Brown <[hidden email]>: > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > I was wondering if there is any clear advantage to structured > illumination versus deconvolution? Can it actually give you higher resolution? > > Sincerely, > > Claire > No virus found in this incoming message. Checked by AVG. Version: 7.5.526 / Virus Database: 270.4.1/1522 - Release Date: 27/06/2008 8:27 AM No virus found in this outgoing message. Checked by AVG. Version: 7.5.526 / Virus Database: 270.4.1/1522 - Release Date: 27/06/2008 8:27 AM |
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In reply to this post by Barbara Foster
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear All Programmable array microscopy looks quite
interesting too,. I believe Cairn are developing a system, primarily as an
alternative to CLSM or Nipkow disc systems. No commercial interest. Darran From: Confocal
Microscopy List [mailto:[hidden email]] On Behalf Of Barbara Foster Dear Listers, Search the CONFOCAL archive at |
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal From now on you can contact me at [hidden email] |
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