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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi all, Does anyone have experience with characterizing PSF's (or other aspects of the optics) at different temperatures? For instance, how much effect is there on an objective when the temp is raised from room temp (25C) to physiologically relevant 37C? Thanks, Carl Carl A. Boswell, Ph.D. Molecular and Cellular Biology University of Arizona 520-954-7053 FAX 520-621-3709 |
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal The difference will come with the SA correction. TIRF lenses (at least the Leica one) actually have values for different temperatures on the correction ring. This is working with oil, which changes RI rather a lot with temperature. In water the difference will be smaller, but it's still worth setting the correction collar visually rather than by trusting the scale. Once you have set it correctly there should be no real difference in PSF. 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 Carl Boswell Sent: Friday, 8 February 2008 5:58 AM To: [hidden email] Subject: heat and PSF Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi all, Does anyone have experience with characterizing PSF's (or other aspects of the optics) at different temperatures? For instance, how much effect is there on an objective when the temp is raised from room temp (25C) to physiologically relevant 37C? Thanks, Carl Carl A. Boswell, Ph.D. Molecular and Cellular Biology University of Arizona 520-954-7053 FAX 520-621-3709 No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.5.516 / Virus Database: 269.19.19/1258 - Release Date: 4/02/2008 10:10 AM No virus found in this outgoing message. Checked by AVG Free Edition. Version: 7.5.516 / Virus Database: 269.19.19/1258 - Release Date: 4/02/2008 10:10 AM |
In reply to this post by Boswell, Carl A - (cboswell)
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal The psf changes not too much in x and y but increased in Z (RT compared to 37oC). So you will lose resolution mostly in z. This was measuresd with fixed tetraspeck beads 63x oil objective. I have no idea how the psf behaves with other objectives or during life imaging. Cooling from 37oC to RT untill the PSF is stable again took on our system about 2.5 to 3 hours.(all done with the meta LSM 510). Mariette ____________________________________________ Dr. M.P.C. Kemner-van de Corput ____________________________________________ MGC - Dept. of Cell Biology & Genetics Erasmus Medical Center Dr. Molewaterplein 50, 3015 GE Rotterdam POB 2040, 3000 CA Rotterdam The Netherlands Office: H-Ee751; Lab: H-Ee710 tel office: +31 10 704.3949 tel lab: +31 10 704.3315 tel secr: +31 10 704.3169 ____________________________________________ http://www2.eur.nl/fgg/ch1/cellbiology/ http://www.thesis.kemner.biz/ ____________________________________________ Op Do, 7 februari, 2008 7:57 pm, schreef Carl Boswell: > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > Hi all, > Does anyone have experience with characterizing PSF's (or other aspects of > the optics) at different temperatures? For instance, how much effect is > there on an objective when the temp is raised from room temp (25C) to > physiologically relevant 37C? > Thanks, > Carl > > Carl A. Boswell, Ph.D. > Molecular and Cellular Biology > University of Arizona > 520-954-7053 > FAX 520-621-3709 > |
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