Dear Muhammad, Assuming you are using the common "Rhodamine" that used to be known as "TRITC" years ago, it will not be excited by a 488 laser so it's not surprising that you could not detect it. Your supplier should be able to provide absorption and emission spectra for the dye you are using to verify this. I think you have two options: 1- Use a different laser wavelength to excite the dye. This of course depends on the flow cytometers and lasers you have access to. 2- Substitute another dye that will be excited by your laser. That may be a completely different dye, or may be a variant of Rhodamine that has a lower excitation wavelength; there are several listed among the spectra on the Molecular Probes web site, no doubt there are many other sources. Hope this helps, Joseph. At 05:25 29/3/2001, Muhammad Anzar wrote: >Hello Friends: >We are working on the uptake of a rhodamine-labeled protein by >spermatozoa. Under fluorescent microscope, there is clear fluorescence >on sperm head but but it can not be detected by flow cytometer (argon >ion laser of 488 nm) using FL3 (wavelength 580). Two different flow >cytometers have been tried but the results are same. Your suggestions in >solving this problem would be highly appreciated. > >Muhammad Anzar DVM, PhD >Department of Animal and Poultry Science >University of Guelph >Guelph, Ontario, Canada N1G 2W1 >Phone: 519-824-4120 (x3689 Off; x8355 Lab) >Fax: 519-836-98733 -- Joseph Webster, Flow Cytometry Facility Centenary Institute, Sydney AUSTRALIA.
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:13 EST