[Histonet] Re: cells in agar

[Brian Chelack]

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.


Brian Chelack
Prairie Diagnostic Services
2604-52 Campus Drive
Saskatoon SK
S7N 5B4
306-966-7241