epithelial cells + flow, newbie question

From: Maciej Simm (simmmmer@yahoo.com)
Date: Tue Dec 21 1999 - 10:43:55 EST


Greetings once again,

first, Happy Holidays everyone! I hope I get all the replies to this 
message before y2k bug chews up all the wiring in our computers :)

issue #1
I have recently been working with a PI who studies epithelial cells. 
During acquisition, my instrument was calibrated using BD calibrite 
beads, 2 color. I noticed that using ssc @ 1.00 and fsc @ 2.00 a lot
of those cells would still not "fit" in the panel. I decreased fsc to
1.00 and then decided to change fsc and ssc modes from log to lin. The
question is, by doing that, am I acquiring more events or less? Is the
coverage of ssc vs. fsc broader in lin or log? If there's a FAQ on
this I would love to read it. 

issue#2 (a little off topic..)
The PI has some problems with epithelial cells' clumping and adhering.
He was told to try trypsin to get them off the plastic tubes, but he
is afraid that might affect his protein of interest which is expressed 
on the surface. Another thing he tried was to soak his cells in PBS
without calcium, but he thinks they tend to clump that way and it's hard
to obtain cell counts. What is the best way to prepare epithelial cells
for flow?


Thanks in advance for any responses. 

=====
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|    Maciej S. Simm      | 525 E 68th Street  |
|  Research Technician   | Room N-805         |
| Cornell Medical Center | Tel. 212.746.3428  |
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