[Histonet] Cell blocks,

[Monfils, Paul]

Here's my standard cell block technique, which I think is pretty simple:


Get the cells into suspension in a centrifuge tube.

Add a volume of 10% buffered formalin equal to the volume of the suspension
(resulting in 5% buffered formalin) and mix by inversion or vortexing.

Allow to fix for 10 minutes or longer.

Add 1 drop of Mayer's albumin per ml of sample and mix by inversion or
vortexing.

Spin down and gently remove supernatant, by pipetting or pouring.

Gently add 50% ethanol (hold the tube at a 45 degree angle and slowly
pipette the alcohol onto the side of the tube, not directly onto the cells).
Allow to stand 10 to 30 minutes, depending on the volume of the cell block.

Pour off, replace with 70% ethanol.  Allow to stand a similar time.

Pour off, replace with 95% ethanol.  Allow to stand a similar time.

Remove cell block from centrifuge tube, wrap gently in lens paper, place in
cassette, process and embed.


The albumin coats the cells with a thin film of coagulable material, which
subsequently coagulates in the alcohols, binding the cells together into a
block.  The 50% alcohol should be added carefully, so as not to cause
turbulence that might bring some cells back into suspension.  The 50%
alcohol coagulates the albumin sufficiently so that the subsequent alcohols
can be added less gently. Other proteinaceous material like gelatin or serum
could substitute for albumin.  The cell block shrinks somewhat during
dehydration, just as tissue does, and usually comes loose from the bottom of
the tube as a result.  If it does not come loose on its own, just touching
it with the tip of a probe will usually free it.


> ----------
> From: 	histonet-bounces <@t> lists.utsouthwestern.edu on behalf of
> Steven Coakley
> Sent: 	Friday, March 10, 2006 1:07 PM
> To: 	Histonet <@t> lists.utsouthwestern.edu
> Subject: 	[Histonet] Cell blocks,
> 
> I'm looking for a way to make simple cell blocks.  I work in research so I
> need something that would cover routine cells and cell cultures??
>    
>   Steve
> 
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