[Histonet] Plasma/thrombin clot for cell block

[Linda Prasad (SCHN)]

Hi Cheryl

This is the technique we use.

Thromboplastin Cell Block method

Principle:

The Thromboplastin cell block is very gentle on cells and allows a wide range of routine immunohistochemical stains to be done.
Plasma, treated with EDTA, is mixed with the specimen in the presence of thromboplastin and calcium ions to form a cell clot.
This method is not suitable for cells fixed in formalin or specimens of bile fluid (bile enzymes prevent clot formation)

Solutions:

1.	Normal Plasma, containing EDTA , use out of date plasma obtained from Blood Bank. Freeze in 2ml aliquots. This plasma has been tested for known viruses and is considered safe.

2.	Thromboplastin, use out of date thromboplastin obtained from Haematology.

3.	1% calcium chloride

4.	10% formalin

Method:

1.	Spin fluid using a centrifuge.
2.	Decant off supernatant
3.	Resuspend pellet in 3 drops of plasma.
4.	Add 3 drops of thromboplastin, mix
5.	Add 3 drops of Calcium Chloride, mix gently
6.	Allow to stand undisturbed for 15-20 minutes
7.	Add 5-8ml 10% formalin and allow to fix
8.	Place in a labelled tissue cassette and process as usual.

Reference: 
Furtado (1970) Stain Technol 45(1):19-23.

Thank you :)

Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.prasad at health.nsw.gov.au | w: www.schn.health.nsw.gov.au


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-----Original Message-----
From: Cheryl [mailto:tkngflght at yahoo.com] 
Sent: Wednesday, 6 May 2015 2:19 AM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] Plasma/thrombin clot for cell block

HI Guys-

I need procedures including products to create a plasma cell block for non-gyn cytology.

The one we have uses bovine thrombin and calcium chloride (dihydrate) but it's gotten corrupted and I can't find the original resources.

Help?

Cheryl
tkngflght at yahoo.com

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