[Histonet] (no subject)
Questions about LCM.
I am attempting a unique application of Laser Capture. In order isolate pure
ovarian surface epithelium I am taking the ovary directly in the OR and
rolling it between two glass slides. The surface epithelium easily detaches
and sticks (loosely) to the slide. I then follow this protocol for staining
slides are frozen in the OR.
5-10 minutes acetone/EtOH fixation (3:1)
graded EtOH 1 minute each
5 minutes xylenes
proceed with LCM
The probem I am having is that the cells sometimes become too firmly
attached to the slide and will not under any means come off the slide with
the LCM scope. I do not have a lot of material to play with so I would
appreciate any help anyone might have. Will a longer xylenes incubation
detach the cells or a shorter fixation? I need the cells attached firmly
enough that they dont come off in the solutions but not so firmly attached
that they wont come off the slide via LCM. Does anyone have experience with
this type of thing?
Thanks for any help.
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