[Cytometry] COL-3

Haviland, David L David.L.Haviland at uth.tmc.edu
Sat Jul 4 22:05:44 EDT 2009


In addition to testing the cells alone for autofluorescence, you may (should) have to find a fluorimeter or scanning spectrophotometer to determine excitation and emission of this drug.   The manufacturer should have this info but it may be easier for you to figure it out empirically.    Prima face, I'd say you are dealing with a compensation issue because some little known feature of this drug.   You're going to have to check the cells which are probably ok but also check the drug by itself and factor COL-3 treated cells into your color scheme.   

Since you didn't state, what sort of machine were you using?  I presume it is possible that you could open a number of parameters and compare the histograms in the untreated cells. 

Tell us more when you can.

David

________________________________________
From: cytometry-bounces at lists.purdue.edu [cytometry-bounces at lists.purdue.edu] On Behalf Of Mona Fares [Mona.Fares at ki.se]
Sent: Saturday, July 04, 2009 3:10 PM
To: cytometry at lists.purdue.edu
Subject: [Cytometry] COL-3

Dear all

I stained cells treated with COL-3 with Annexin V and PI, The cells are
shifted to the right so, I cann't determine the positive stained cells
accurately. the non stained treated cells also shifted to the right this
is not the case with the control and with cells treated with other drug.
The drug has a yellow color but I don't know if this could affect the
compensation, also I don't know if the treated cells has some degree of
autofluorescence.
Does any one worked with COL-3 before or any one has any suggestion for
instrument setting and analysis?.

Mona Fares
Research fellow
Karolinska Institute
Sweden
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