[Histonet] Re: cells in agar
brian.chelack <@t> usask.ca
Mon Jan 16 16:38:26 CST 2006
I have had good success with a bit simpler method of creating a cell pellet
that can be subsequently fixed, sectioned and immunostained. Melt a small
volume of 2% low melting point agarose in normal saline. Hold at 37C.
Re-suspend the cells of interest at about 1 10e6/ml and mix (using a Pasteur
pipette) with an equal quantity of the melted agarose. Now to make real nice
little agarose plugs I quickly add the still melted mixture into wells of a
96 well polyethylene plate (one of the fexible ones) and place it in the
fridge for a few minutes. After the agarose/cell mixture is set, pop out the
agarose plugs and place directly into 10% NBF. After fixation, the plugs can
be transected and processed into paraffin blocks. If you don't have a
polyethylene 96 well plate you can just drip the mixture onto a glass slide
and peel off the drops after they have set.
A bit low tech, but it works for me.
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