[Histonet] frozen section problem

[Manfre, Philip]

Hi Dorianne,
	You need to freeze your tissue faster.  Ideally, isopentane placed in a metal cup, that is that is then frozen in a dewar of liquid nitrogen, works best.  The isopentane, once frozen, is thawed a little with a metal rod to produce a small liquid pool and your tissue is placed in this for about one minute.  You need some equipment for this procedure, such as the metal cup that can sit inside a small, open dewar of liquid nitrogen.  Alternatively, you can freeze directly in liquid nitrogen, though you need to beware of the tissue fracturing due to the sudden and extreme temperature reduction.  Slower freezing of tissue (sitting on dry ice, etc.)  allows ice crystals to form in the tissue, creating the vacuoles you describe.
	I hope this helps.
Phil.

Philip Manfre, BA, HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP81-406
770 Sumneytown Pike
West Point, PA 19486
215-652-9750
philip_manfre at merck.com




-----Original Message-----
From: Bonello Dorianne M at Health-MDH via Histonet <histonet at lists.utsouthwestern.edu> 
Sent: Friday, July 16, 2021 11:25 AM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] frozen section problem

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Dear all,


We are experiencing freezing artifacts on our frozen sections. Basically, we are seeing cavity-like structures under the microscope, mostly elongated, especially when it's a frozen section on brain tissue. This is most probably happening due to ice crystal formation. We're not using cryospray, relying only on the cryobar boost function.


Does anyone has a solution to this problem please?


Regards,



Dorianne Bonello
Allied Health Practitioner (MLS)
Histology Laboratory - Pathology
Health-Mater Dei Hospital


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T +356 +356 25456434

E dorianne.m.bonello at gov.mt


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