Flow on Nematode eggs

McCoy, J. Philip (NHLBI) McCoyJ at nhlbi.nih.gov
Fri Mar 1 08:05:01 EST 2002


 Hi Janet;

Just yesterday we sorted newly hatched C elegans. We used a 200 micron tip,
and of course, low pressure. We found forward scatter to be of limited use
in analyzing these worms - possibly due to their varied orientation in the
stream at the low pressure. AS always, I would check the specimen under a
microscope for doublets, staining, etc before sorting.


Phil


-----Original Message-----
From: janet dow
To: cyto-inbox
Sent: 2/28/2002 10:26 AM
Subject: Flow on Nematode eggs


I have a client who wish to run flow on nematode eggs (Meloidigyne
incognita and Meloidigyne hapla) to stain to look at genome size.  We
have
tried twice to run them and the first time saw no florescence so we
change
protocols to PI but the second time we clogged both my machines with the
samples( I am using a BD FACScan and a FACSCalibur).   Can anyone help
us-she has found a paper from 1984 which used flow on a machine made at
Los
Alamos but it doesn't specify what size nozzle they used.  She also does
not know how big the eggs are but you can visualize them in the solution
and they settle out of solution very quickly.


thank you in advance for all your help-I have always had such great luck
with questions on this group.

Sincerely

Janet Dow


Janet Dow
Research Technician and Manager
Flow Cytometry Facility
North Carolina State College of Veterinary Medicine
Room C-314
Raleigh, NC 27606
(919)513-6364




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