[Histonet] Cell block preparations

[Tony Henwood (SCHN)]

Hi Jennifer,

I have had excellent success with lysing the red blood cells (using  Isotonic Ammonium Chloride) prior to cell block preparation with thromboplastin-plasma. 
The lysing solution contains EDTA so you will need to add a few drops of 1% calcium chloride. Method as follows:

Lysis solution
Ammonium Chloride        4.5g
Potassium carbonate      0.5g
EDTA                     0.0186g
Distilled water          500mls

Method:
1.	Centrifuge bloody fluid.
2.	Remove supernatant and add equal volume of lysis solution.
3.	Resuspend and incubate for 5 minutes at 4oC.
4.	Centrifuge, if blood still remains, then repeat from step 2.
5.     Rinse in Hanks or RPMI, centrifuge.    
6.	Mix pellet in a few drops of plasma.
7.     Add thromboplastin and a few drops  of 1% Calcium Chloride, mix gently and allow clot to form.
8.     Add 10% buffered formalin and fix and process as usual.

Reference:  
Kuenen-Boumiester etal (1996) Acta Cytolog 40:475-479

If you donot use the plasma clot method for cell block preparation, then use your preferred method after step 5.
The lysis solution can also be purchased commercially from several companies (eg Biolegend). It is commonly used for sample preparation for flow cytometry. Check the SDS to make sure it does not contain formaldehyde.


-----Original Message-----
From: Mac Donald, Jennifer via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Friday, 17 January 2020 4:53 AM
To: Charles Riley <criley at dpspa.com>; Histo List <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Cell block preparations

Acetic acid would work.

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From: Charles Riley via Histonet <histonet at lists.utsouthwestern.edu>
Sent: Thursday, January 16, 2020 8:55:21 AM
To: Histo List <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Cell block preparations

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What is the best way to remove excess blood from FNA sample collections before spinning them down into cell blocks?

--

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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