Re: Embedding cells

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From:"Joyce Kotzuk" <>
To:<>, <>
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I was given "some cells" recently in eppendorf tubes and asked to stain and put them on slides to visualize under light microscope. As I have never worked with cells before, I had no idea where to start. I was told to begin by "spinning them down", but at what speed and for how long? When I did, at the rather slow speed of 1000 rpm for 5 minutes, (I was concerned about damaging the cells), there was no pellet or anything visible to my eye at all. I tried all manner of fixing with ethanol, acetone, staining with cresyl violet or hematoxylin, pipetting a drop onto a slide and letting it dry, then staining, all to no avail. I never saw any cells anywhere. It was the most frustrating day I've had in a long time. Anyway, this seems related to the question asked here, and I would love to know as well, how to treat cells for fixing, staining and visualizing.
Thanks in advance, Joyce Kotzuk, UNM pathology

>>> Emma Carter <> 04/26/00 09:40AM >>>

I am sure i read a message about this recently, but i cant recall, and i
dont have anything in my mail folder...

someone has asked me about paraffin embedding some cells. I came up with a
protocol off the top of my head, roughly:

Put cells in eppendorf tube, spin down and remove culture buffer.
Resuspend with formalin and leave for 10 mins. Spin down and remove
Resuspend/spin down through serial alcohols, then xylene.
after final resuspension with xylene, remove and cover with warm wax.
Allow wax pellet to set. Pop out of tube and embed in cassette.

How does this appear to other people? Should i go through the dehydration

Any help would be very appreciated...... (oh yeah, this is all to be done by
my own fair hand.......automation!? Pah!)

Thanks in advance,

emma carter

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