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
More information about the Cellbiol