Help for the TUNEL method for apoptosis

Immunobiology Research Institute, Siena (unutmaz@iris02.biocine.it)
Tue, 2 May 1995 19:03:49 +0200

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Recently we have been trying to setup the TUNEL method to detect apoptosis
of mouse thymocytes in suspension, according to the published method by Kishimoto H.
et. al. 1995 J. Exp. Med. 181:649. But we have a major problem, that is the recovery
of cells, expecially after refixation with 1% paraformaldehyde following initial
fixation with 70% ethanol. At this step we lose more than 95% of our starting cells!

So far we have tried different spinning conditions but the yield never improved. Is
there a particular speed and/or temperature to centrifuge the cells after the
paraformaldehyde treatment?

I would very much appreciated any suggestions.
Thanks in advance.

Derya,
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Derya Unutmaz,
Immunobiology Research Institute,
Siena, Italy.
E-mail: Unutmaz@iris02.biocine.it
Fax: +39-577-24 35 64

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