Formaldehyde used on smears does alter the formation of the purple
Azur-Eosin staining complex. Dipping the smears in methanol, Bouin's or even
Susa's solution before staining may improve your results. Stock solutions
stored in containers with metal lids for any period of time should also be
avoided. It is also possible that the organic solvent in your stock solution
may have evaporated causing subsequent precipitation.

Troubleshooting Romanowsky-Giemsa staining is explained in detail by Horobin
& Walters (1987) and Wittekind & Gehring (1985).



Eric C. Kellar
University of Pittsburgh Medical Center


	----------
	From:  Gayle Callis [SMTP:uvsgc@msu.oscs.montana.edu]
	Sent:  Sunday, March 05, 2000 7:18 AM
	To:  histonet@pathology.swmed.edu
	Subject:  chromosome staining

	Something I thought I would never have to do, alas, I need the quick
Giemsa
	method for chromosome preps.  All he wants to do is see the
chromosomes
	without banding, and has used the standard fixation to release the
'somes
	from the cell, then splash then onto the slide.  He loses them
unless he
	flames, but the Giemsa we did, on his recommendation, is stain,
rinse, dry
	and coverslip.  Originally he just dipped in the stain, and my book
said a
	Giemsa stain would work, but no specific technic, that may be where
things
	went cockeyed.  

	Ppt everywhere, after flooding the slide with Giemsa for 6 min, then
	rinsing off, drying.  May be the stain solution is too old, and
filtering
	is necessary, the next thing.  

	Is there a better way, brief, quick and dirty.  My Giemsa is the
Harleco aka
	EM Diagnostic Systems original azure a blend, not the newest lot.
Are we
	doing something wrong in the beginning.  He had used the Sigma
Giemsa
	previously, and said the chromosomes were blue, now they look more
like
	light purple.  

	Since this is not my area of expertise, I am groping in the dark.
My new
	Humason's book was helpful, but something is missing.






	Gayle Callis
	Veterinary Molecular Biology
	Montana State University
	Bozeman MT 59717-3610
	406 994-4705
	406 994-4303