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