In my opinion the realtime autofocus solutions offered today are
absolutely essential, they are incredibly useful for live cell imaging and
allow multiday non-attended timelapse. However for the system to
work properly there generally has to be a significant refractive index change
from the material being imaged (for example from the coverslip 1.51 to media,
1.36). I have never used a plastic substrate( such as a 96 well
plate) on a focussing system, I would be surprised if they work at all. so
make sure this is possible before investing, perhaps some of our commercial
friends can comment on the utility of real time focussing systems for use on
plastic
S
Simon C. Watkins Ph.D, FRCPath
Professor and Vice Chair, Cell Biology and Physiology
Professor, Immunology
Director, Center for Biologic Imaging
BSTS 225, University of Pittsburgh
3500 Terrace St.
Pittsburgh PA 15261
Tel: 412-352-2277
Fax:412-648-2797
URL: http://www.cbi.pitt.edu
From: Confocal Microscopy
List [mailto:[hidden email]] On Behalf Of Mayandi
Sivaguru
Sent: Thursday, September 18, 2008 10:10 AM
To: [hidden email]
Subject: Re: Perfect Focus
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Scott, it looks like you have a fully automated system. Personally, I do not
rely on autofocus, I would suggest, if you take multi position, multi channels
over multiple time point, then instead of one frame position/well also include
a small z stack, say 4 images (2 above and 2 below) from the best focus of each
position, so latter even if there is a drift, you will have an option to select
the best plane out of those four.
Shiv
At 07:56 AM 9/18/2008, you wrote:
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
All,
Have new faculty coming aboard and a major application will be 48 hour
timelapse studies in 96 well plates using multichannel collections.
Therefore my question is how critical is it to have a system that has
a perfect focus mechanism to meet this application? Are there decent
work arounds if the perfect focus is not used? Have never actually
worked with this type of system so would like input on this matter
from the list. Thanks.
Scott J. Howell, Ph.D.
Manager, Imaging Module
Visual Sciences Research Center
Case Western Reserve University
2085 Adelbert Rd.
Institute of Pathology Room 106
Cleveland, Ohio 44106
216-368-2300
http://www.case.edu/med/vsrc/
Mayandi Sivaguru, PhD, PhD
Microscopy Facility Manager
8, Institute for Genomic Biology
University of Illinois at Urbana-Champaign
1206 West Gregory Dr.
Urbana, IL 61801 USA
Office: 217.333.1214
Fax: 217.244.2496
[hidden email]
http://core.igb.uiuc.edu
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