Current Protocol for Metphase Chromosomes?

John Ladasky (ladasky@leland.Stanford.EDU)
Mon, 27 Nov 1995 15:49:57 -0800 (PST)

Greetings, everyone,

I'm interested in performing one-dimensional flow karyotyping of
PHA-stimulated monkey leukocytes or EBV cell lines, using PI as the stain.
I currently have two references for the preparation of metaphase chromosomes:

Lundsteen C et. al., Cytometry 19:42 [1995]

Aten JA et. al., Histochemistry 87:359 [1987]

The first article uses colcemid to effect a metaphase block, but
the concentration is not specified. The older article uses a blocking
reagent I've never encountered, called vindesine. Is vindesine obsolete?
At least they tell me what concentration they used.

Do you have a favorite protocol?

Is there a comprehensive discussion of this technology available
anywhere -- say, a recent review or book chapter? Can I expect EBV-trans-
formed cell lines to show chromosomal alterations relative to the parental
cells?

Thanks in advance,
- John Ladasky

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