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 Normal Plasma, containing EDTA , use out of date plasma obtained from Blood Bank.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 Thromboplastin, use out of date thromboplastin obtained from Haematology. 3. 1% calcium chloride 4. 10% formalin Method: 1.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: [email protected] | w: www.schn.health.nsw.gov.au Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia Locked Bag 4001, Westmead 2145, NSW Australia ♲ Please consider the environment before printing this email. -----Original Message----- From: Cheryl [mailto:[email protected]] Sent: Wednesday, 6 May 2015 2:19 AM To: [email protected] 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 [email protected] _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************************* This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Sydney Children's Hospitals Network. This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Sydney Childrens Hospital's Network accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************************* _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
