Flowcytometry of testicular cells

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msvnathan
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Flowcytometry of testicular cells

hi
i did flowcytometry for cells isolated from testis. I used propidium iodide in the concentration of 50 microgram per ml. But i am facing problem while going for flowcytometry. They said concentration of PI is low so signal for fluro is not appearing. is this concentration is ok or concentration should be changed please mention.  

Arvind Singh Pundir
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refer the FACS analysis in the below mentioned paper in which they have succesfully used .05mg/ml of PI, generally 5 microgram/ml  of PI works well

"Developmental Schedule of the Postnatal Rat Testis Determined by Flow Cytometry"
www.biolreprod.org/cgi/reprint/59/1/84.pdf

msvnathan
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hi Pundir,

hi Pundir,
Thanks for the paper. i have read this paper, they also used 5 microgram per ml only. i will try according to this paper this time and reply. Thanks once again.

Jason King
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Are the cells you are FACS

Are the cells you are FACS analysing fixed?

msvnathan
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Dear Mr. Parvoman,

Dear Mr. Parvoman,
Yes my samples after separation from testis i will fix it in 70% methanol for 24hrs then PI is added and incuabated for 40 min.