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From:Tamara Howard <>, Histology listserver <>
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DAPI staining is a piece of cake - incubate the fixed cells in the
solution for 5-10 minutes. Wash several times in buffer before mounting
for microscopy. Some people put the DAPI in the mounting medium but I
think the background with that method is horrendous. As for the dilution
of the DAPI - anything from 1mg/ml down to 1 nanogram/ml! You are supposed
to be able to see Barr bodies if you get the DAPI diluted out far enough,
but I've never had the DAPI so dilute that everything else dropped out.
Oh, and DAPI only works on fixed (permeabilized) cells. 

You'll need to run some non-apoptotics as a control - DAPI looks different
in different cell types. For apops, you should see staining around the
nuclear margins and blobs within the nucleoplasm. But mouse cells always
look blobby by DAPI.....

Good luck!

Tamara Howard

>Does anyone have any experience staining with DAPI?  I was told that DAPI
>staining would be sufficient for demonstration of apoptosis.  I have
>cells on coverslips and am trying to find the least complex method. 
>Apotag seems cumbersome and I would love something with fewer steps.
>Thanks in advance!
>Nancy Lemke
>Henry Ford Hospital
>Neurosurgery Research

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