Re: chromosome staining

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From:rkline@emscience.com
To:Gayle Callis <uvsgc@msu.oscs.montana.edu>
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Gayle,

There is a Giemsa Stain specific for G-banding from Harleco.  The catalog number
is 620G/75.  I can send you a procedure that was used by one of the labs that
did our QC testing.

You need new Giemsa.

Also, the pH of the buffer is very important.  It controls the intensity ( among
other things) of the stain.

You know where to find me.

Regards,
Rande Kline, HT (ASCP)
Technical Services
EM Science/BDH






Gayle Callis <uvsgc@msu.oscs.montana.edu> on 03/05/2000 07:17:30 AM

To:   histonet@pathology.swmed.edu
cc:    (bcc: Rande Kline/EMI/Merck)
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








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