Re: [Histonet] frozen section problem

2021-07-16 Thread Tony Henwood (SCHN) via Histonet 
We have had this issue previously.

We tracked it down to the brain biopsies arriving in "Isotonic" saline (which 
is really not isotonic).
See: Henwood, A., (2007) “Adverse effect of saline on brain intraoperative 
(frozen section) Histology” J Histotechnol 30(3):193.

Ask the surgeons to send the biopsies to the lab on damp shiny (non-absorbent) 
card.

Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Principal Scientist, the Children’s Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney
Tel: 612 9845 3306
Fax: 612 9845 3318
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA


From: Manfre, Philip via Histonet 
Sent: Saturday, 17 July 2021 07:54
To: Bonello Dorianne M at Health-MDH;   (histonet@lists.utsouthwestern.edu)
Subject: Re: [Histonet] frozen section problem

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_man...@merck.com




-Original Message-
From: Bonello Dorianne M at Health-MDH via Histonet 

Sent: Friday, July 16, 2021 11:25 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] frozen section problem

EXTERNAL EMAIL – Use caution with any links or file attachments.

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


[cid:image001.jpg@01D67184.63288530]


T +356 +356 25456434

E dorianne.m.bone...@gov.mt


Mater Dei Hospital, Triq id-Donaturi tad-Demm, l-Imsida, Malta MSD 2090 | Tel 
+356 2545  | 
https://secure-web.cisco.com/108H5-f5Ylbr6vXriY71-fp4JDQxST7FvP8i3GNqBPS05qZlvu_1O6tMXRjgYucUESUfrxOSGo-3VSj277AHeUKp72a2u23a8E1gThp8T_C_qgaCkXVHMnAxY-YyXqqjb0pEmmV-J74L1UJXhwqdO88nHIMcK0Y7_wGmWPWFgvvMfmer4a-NFgWAPDQDYgBdnmJ-UysbSbmKuEVVZzOZFDBgn-f2wXW3JKjvg0h2q9rXdcxCMqWYFuQFY1xiKWJHR9NcNUphJ6xHBSRJSogoK53iKRSk1tEyOBPErMdipZOz4PXDp29G4cAm7LIrI_3TQWhJLOaI2wZTc0wJ7yRUScq8nh2P3hrxSuBaL3ivOfsU9eUtNGDZDo9XeNFEcGmivY-fPo1XKWjQy6pmHUaksuix_Jp9AvaCBh0IWf_zuPVGEv2oeS9aYQXub1b9B0QpJFqSicysNAHY65JYUzhjHXxuuvsaJ-m6zou-55OWaT8dK3Cjh6bvXTnQWzS3C4Uya/https%3A%2F%2Fdeputyprimeminister.gov.mt%2Fen%2FMDH%2FPages%2FHome.aspx<https://secure-web.cisco.com/108H5-f5Ylbr6vXriY71-fp4JDQxST7FvP8i3GNqBPS05qZlvu_1O6tMXRjgYucUESUfrxOSGo-3VSj277AHeUKp72a2u23a8E1gThp8T_C_qgaCkXVHMnAxY-YyXqqjb0pEmmV-J74L1UJXhwqdO88nHIMcK0Y7_wGmWPWFgvvMfmer4a-NFgWAPDQDYgBdnmJ-UysbSbmKuEVVZzOZFDBgn-f2wXW3JKjvg0h2q9rXdcxCMqWYFuQFY1xiKWJHR9NcNUphJ6xHBSRJSogoK53iKRSk1tEyOBPErMdipZOz4PXDp29G4cAm7LIrI_3TQWhJLOaI2wZTc0wJ7yRUScq8nh2P3hrxSuBaL3ivOfsU9eUtNGDZDo9XeNFEcGmivY-fPo1XKWjQy6pmHUaksuix_Jp9AvaCBh0IWf_zuPVGEv2oeS9aYQXub1b9B0QpJFqSicysNAHY65JYUzhjHXxuuvsaJ-m6zou-55OWaT8dK3Cjh6bvXTnQWzS3C4Uya/https%3A%2F%2Fdeputyprimeminister.gov.mt%2Fen%2FMDH%2FPages%2FHome.aspx>
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Re: [Histonet] frozen section problem

2021-07-16 Thread Manfre, Philip via Histonet 
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_man...@merck.com




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

EXTERNAL EMAIL – Use caution with any links or file attachments.

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


[cid:image001.jpg@01D67184.63288530]


T +356 +356 25456434

E dorianne.m.bone...@gov.mt


Mater Dei Hospital, Triq id-Donaturi tad-Demm, l-Imsida, Malta MSD 2090 | Tel 
+356 2545  | 
https://deputyprimeminister.gov.mt/en/MDH/Pages/Home.aspx<https://deputyprimeminister.gov.mt/en/MDH/Pages/Home.aspx>
 | https://www.facebook.com/materdeihospital/


Think before you print. Kindly consider your environmental responsibility.

This email and any files transmitted with it are confidential, may be legally 
privileged and intended solely

for the use of the individual or entity to whom they are addressed.






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Re: [Histonet] frozen section problem

2021-07-16 Thread John Kiernan via Histonet 
Yes, definitely ice crystal holes! If the tissues are unfixed you will have to 
freeze much more rapidly (isopentane cooled with liquid nitrogen.) If fixed in 
formaldehyde, cryoprotect by immersing the pieces in 20% sucrose, until they 
sink.

John Kiernan
Anatomy & Cell Biology, UWO
London, Canada
= = =

From: Bonello Dorianne M at Health-MDH via Histonet 

Sent: July 16, 2021 11:25 AM
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] frozen section problem

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


[cid:image001.jpg@01D67184.63288530]


T +356 +356 25456434

E dorianne.m.bone...@gov.mt


Mater Dei Hospital, Triq id-Donaturi tad-Demm, l-Imsida, Malta MSD 2090 | Tel 
+356 2545  | 
https://deputyprimeminister.gov.mt/en/MDH/Pages/Home.aspx<https://deputyprimeminister.gov.mt/en/MDH/Pages/Home.aspx>
 | https://www.facebook.com/materdeihospital/


Think before you print. Kindly consider your environmental responsibility.

This email and any files transmitted with it are confidential, may be legally 
privileged and intended solely

for the use of the individual or entity to whom they are addressed.






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Re: [Histonet] Frozen section problem

2021-07-16 Thread Bob Richmond via Histonet 
Dorianne Bonello, Allied Health Practitioner (MLS), Histology Laboratory -
Pathology Health-Mater Dei Hospital, on the island of Malta asks:


> >>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.<<
>
> You need to freeze faster. A low-tech solution is a "heat extractor" - a
> flat-bottomed mass of metal with a handle, kept cold in the cryostat, that
> you press on top of the specimen after it's mounted on the chuck with
> frozen section embedding medium. Here's a picture of it:

https://tinyurl.com/4djk85xd

This 82 year old pathologist would expect to trouble-shoot the problem
himself.

Not every day I get e-mail from Malta! If anyone on Histonet doesn't know
where Malta is, it's an island nation in the Mediterranean Sea, south of
Sicily. If you look at the Mater Dei web site, you'll see that it's in
English, but with occasional lines of Maltese, the native language of the
island, a form of North African Arabic, but always written in the Roman
alphabet. Wikipedia has good resources for the island, the language, and
the Mater Dei Hospital.

Bob Richmond
Samurai Pathologist
Maryville, Tennessee, USA
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[Histonet] frozen section problem

2021-07-16 Thread Bonello Dorianne M at Health-MDH via Histonet 
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


[cid:image001.jpg@01D67184.63288530]


T +356 +356 25456434

E dorianne.m.bone...@gov.mt


Mater Dei Hospital, Triq id-Donaturi tad-Demm, l-Imsida, Malta MSD 2090 | Tel 
+356 2545  | 
https://deputyprimeminister.gov.mt/en/MDH/Pages/Home.aspx
 | https://www.facebook.com/materdeihospital/


Think before you print. Kindly consider your environmental responsibility.

This email and any files transmitted with it are confidential, may be legally 
privileged and intended solely

for the use of the individual or entity to whom they are addressed.






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