If you can see the eggs they're at least 100 micrometers, perhaps bigger. Particle size should be at most 50% of orifice, so they may be compatible with the standard cuvette in BD benchtop instruments. Also, keeping concentration to about a million per mL should work. If there is breakup of the eggs, free DNA can make a mess. If you have good staining the eggs should be bright in a fluorescence microscope (blue excitation; orange to red emission)--always a good way see labeling problems. In addition, you can estimate size at low magnification. Dave ======================================== David M. Coder, Ph.D., Consultant in Cytometry email: d_coder@msn.com or dcoder1@hotmail.com tel./messages: 206-499-3446 ----- Original Message ----- From: "janet dow" <jldow@unity.ncsu.edu> To: cyto-inbox Sent: Thursday, February 28, 2002 7:26 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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