nuclear stain

J. A. Kiernan jkiernan at julian.uwo.ca
Wed Aug 30 14:51:16 EST 1995


In article <421s3v$eu4 at oban.cc.ic.ac.uk> "M.Patel" <M.Patel at ic.ac.uk> writes:
>I am looking for a simple histological stain to show nuclei of 
>fixed cultured cells.  I can think of a fluorescent stain, Hoechst 33258, 
>but would prefer to use an ordinary light microscope.

The easiest is toluidine blue, 0.2% in approx. 1% acetic acid to
give a pH of about 4.  Fix the cells by immersion for a few minutes
in 95% ethanol-5% acetic acid. Rinse in 70% alcohol, then in water.
Stain for a couple of minutes.  Wash in water and CHECK WITH A
MICROSCOPE.  Rinse in 1% acetic acid if overstained, then wash again 
in water. If the substrate is plastic, apply a coverslip, using a
neutral aqueous mounting medium.  Otherwise, air-dry thoroughly and
apply a coverslip with a resinous mounting medium.

                                     John A. Kiernan
                                     Department of Anatomy
                                     Univ. of Western Ontario
                                     LONDON, Canada  N6A 5C1
                                     e-mail: kiernan at uwo.ca



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