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The index of refraction of the material and media is critical. I was only
commenting on the thickness.
-Michael
At 02:02 PM 01/12/05 -0800, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Evidently the correct coverslip is critical for TIRF. I would like to
>hear the answer to this from Olympus, Zeiss, etc.
>
>Tom
>
>-----Original Message-----
>From: Michael Cammer [mailto:[log in to unmask]]
>Sent: Wednesday, January 12, 2005 1:57 PM
>To: [log in to unmask]
>Subject: Re: Cover slip thickness - P.S.!
>
>
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>And what about TIRF? Does this matter?
>After all, chromatic aberration shouldn't be much of a problem since only a
>narrow range (e.g. 515-560nm) is being imaged at any given time and a depth
>of 200 nm or less means that as long as you can get TIRF, then the RI of
>the media shouldn't matter from the point of view of scrambling the light.
>Just for the sake of splitting hairs and joining the pariah club...
>-Michael
>
> >At the risk of becoming a pariah on the list, I will respectfully disagree.
> >
> >Unless you're either (a) looking at cells growing on coverslips, or (b)
> >a hell of a lot better at mounting coverslips than I am, a #1.5
> >coverglass is NOT NECESSARILY optimal. The mounting medium between the
> >coverslip and the specimen is also part of the optics. If you have,
> >say, 50 microns of DPX or Entallin beneath your coverslip, you might
> >rather use something thinner than a #1.5. I typically use a #1 and
> >still have to adjust my correction collar toward the thick end. But
> >then I really slather on the mounting medium.
> >
> >As always, it's the details....
> >
> >Martin Wessendorf
> >
> >--
> >Martin Wessendorf, Ph.D. office: (612) 626-0145
> >Assoc Prof, Dept Neuroscience lab: (612) 624-2991
> >University of Minnesota Preferred FAX: (612) 624-8118
> >6-145 Jackson Hall, 321 Church St. SE Dept Fax: (612) 626-5009
> >Minneapolis, MN 55455 E-mail: [log in to unmask]
>
>____________________________________________________________________________
>Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med.
>Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461
>(718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
> **This electronic transmission contains information that is privileged.**
____________________________________________________________________________
Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med.
Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461
(718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
**This electronic transmission contains information that is privileged.**
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