Hello,
I would be VERY interested in any responses you might get! We are
struggling with sorting stem cells onto slides and keeping them down during
a subsequent FISH labeling procedure...thanks in advance.
-Bill Hyun
----My original question:
>what is your preferred technique for keeping cells stuck to glass slides,
>post sorting, so as to permit subsequent fixing and morphological analysis
>by cytochemistry? Any technique that works from sorted droplet DIRECTLY to
>slide (that gets rid of saline), for those situations where there are too
>few sorted cells to reanalyze/ make cytosmear with?
> Does CELL-TAk , SUPER FROST , or some other sticky stuff work?
>thanks.
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response #1
When sorting onto slides we use very dilute saline (10 ml of Isoton in 2
litres of distilled water). We then dry at 50 degrees. We do not coat
slides.
Cells adhere well. Droplet deflection is just as good.
Good luck.
_______________________________________________
Graham Vesey
Australian Environmental Flow Cytometry Group
School of Biological Sciences, .-.--:_:\
Macquarie University, _/ \
Sydney, : AEFCG |
Australia NSW 2109. \_ /
Tel- 612 850 8150 '-''''\__/
Fax- 612 850 8174 V
_______________________________________________
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response #2
Lan Bo Chen, around the corner from you at the Farber has some
magical slides that he gets from Germany. The slides are treated by
a proprietary method. He claims to retain >90%
of the cells through rigorous processing.
Albert D Donnenberg, PhD
University of Pittsburgh
Albert D. Donnenberg, PhD
Associate Professor of Medicine
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response #3
Centrifugal Cytology works. I haven built special buckets. The slides
should be clean and then coated with polylysine or the best binding will be
with Meyer's Albumin fixative. The trick is to fix the cells while the
centrifugal field is applied. Glyoxal is a good fixative.
From: Robert C. Leif, Ph.D., PMIAC,
Vice President & Research Director
Newport Instruments
5648 Toyon Road
San Diego, CA 92115
Tel. & Fax (619) 582-0437
E-Mail rleif@crash.cts.com
I will add any future responses- thanks!
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Frederic I. Preffer
preffer@helix.mgh.harvard.edu
Department of Pathology- Cox5
100 Blossom St
Massachusetts General Hospital
Boston MA 02114
(617) 726-7481 {fax x2365}
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