Acridine Orange and Sperm Chromatin Structure

From: Rahul Rathi (rathiresearch@yahoo.com)
Date: Fri Oct 11 2002 - 16:39:11 EST


Dear List members,







We are planning to work with Sperm Chromatin Structure Assay using Acridine Orange. I
have some queries for those who have worked in this area and have published papers,

1) When you talk about red fluorescence does that mean FL-3 values? and the same for
the Green fluorescence meaning the FL-1 values?



2) So, does the Total Fluorescence mean FL1 values + FL3 values?



3) I got hold of the raw data from a FITC-PSA and EthD experiment. When I asked the
machine (computer with FACS) to calculate FL-1 + FL-3 (Probably the Total fluorescence)
values for me and then FL-3/FL1+FL3 (Probably the alpha-t), it gave me totally different
values then what I calculated using the EXCEL. So I got confused as to what exactly do
we mean by total fluorescence, is it FL1+FL3 or does it take into consideration some
other parameters too. However, the computer calculated and self (excel) calculated
Total fluorescence and alpha-t values were highly correlated.



I hope I am clear in my questions.



Rahul




Rahul Rathi BVSc, MS, PhD
Dept of Clinical Sciences,
Faculty of Veterinary Medicine,    Tel : 785-532-4413/5700
Kansas State University,             Fax: 785-532-4989
Manhattan, 66505, KS                 rrathi@vet.ksu.edu
USA
"Research is to see what everybody else has seen, and to think what nobody else has
thought"- Albert Szent-Gyorgyi


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