Re: Perfect Focus

Posted by Mancini, Michael A on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Perfect-Focus-tp1098084p1098428.html

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Yes, I concur, the commercially-available automated scopes can focus fairly well on plastic plates, and the plates themselves are varyingly “flat.”  However, not all of these screening scopes can focus equally-well, not by a long shot. Depending upon the system, they can focus on the substratum, or the cells themselves; some can focus fine at low mag, only, and some can go all the way to 100x (with varying time it takes to reach focus---an increasingly important issue as the number of wells to be imaged increases.  

For an academic core facility that is moving into automated microscopy, scopes that can focus on higher mag/higher NA lenses are likely the most useful/versatile; if you will never “screen” at 40x or higher, then the systems that only go to 20x, or a low NA 40x, should be fine.  Just depends what you need.....  

And here is a quick head’s up for anyone using glass or plastic bottom multi-well plate: keep in mind some of the vendors put out plates that have considerable amounts of intrinsic estrogenic activity (from the various plasticizers (e.g., bisphenol A, etc) and/or glue used to manufacture the dish, or adfix the bottoms)......    Some endocrine studies could be considerably influenced by such “noise” in the system.  

Mike





On 9/18/08 9:51 AM, "Gabor Csucs" <[hidden email]> wrote:

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Answering to the question of Simon:
I can't comment on all the auto-focus solutions, but many of them
(especially the ones on the specialized screening microscopes) do work
with plastic bottom plates.
So we do a lot of AF based imaging in plastic bottom plates (different
brands) using air objectives.
Nevertheless the companies don't guarantee that the AF devices work
equally well with all objectives.

Cheers   Gabor

--
Gabor Csucs
Light Microscopy Centre, ETH Zurich
Schafmattstrasse 18, HPM F16
CH-8093, Zurich, Switzerland

Web: www.lmc.ethz.ch
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