[Histonet] DAPI staining for apoptotic nuclei in frozen sections

From:Guillermo Palao

Hello all,
  I am trying to detect apoptotic nuclei by DAPI staining as a morphological confirmation of FITC-TUNEL positive cells in frozen sections. I stain with DAPI 0.5 ug/ml in PBS for 5 min RT, wash in PBS once, mounting medium and cover. The problem is my DAPI signal is very strong, all nuclei look almost white instead of blue, also nucleoli do not look like "dark spots" in nuclei as in published images, but as very bright dense small white dots. Is it just a question of DAPI concentration? If so, how much could I decrease it? Most articles employ 1 ug/ml for apoptosis detection so I am concerned I might be doing something wrong. Any help is really appreciated.

Guillermo Palao, MD. Ph.D.
Laboratorio de Reumatología
Centro de Investigación
Hospital 12 de Octubre
Avda de Córdoba s/n
Madrid 28041

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