[Histonet] Freezing human skin biopsies for cytokine ICC

[patsy ruegg]

Jamie,
I would ask that the samples be surrounded (cryoprotected) in OCT, then snap
frozen and then sent to you on dry ice.
Peprotech may have some of the cytokines that may work even on ffpe tissues,
on frozens for sure.  We are helping them to optimize some of their
antibodies usually used in Elisa/westerns to tissue.
Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. #216
Aurora, CO 80010
720-859-4060
fax 720-859-4110
pruegg <@t> ihctech.net
www.ihctech.net 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gudrun Lang
Sent: Saturday, February 17, 2007 2:58 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: AW: [Histonet] Freezing human skin biopsies for cytokine ICC

First I have to admit that I have no idea how sensible these cytokines are.
We do immunofluorescence on skin biopsies (immunoglobulins, fibrinogen). The
skin biopsies are sent to us in an humid chamber (a tube with NaCl-humid
gaze, not swimming) within 30 min after excision. 
We orientate and freeze them like other tissue, where frozen cuts are
required: put them on the chuck, surround with OCT and let freeze at -20
degree in the cryocut. Afterwards we put two layers parafilm around the
frozen chuck and store them in the refrigerator -20°C until we do the IF
(within a few days). 
You can collect the OCT-blocks and transport them in a cryobox with dry ice.
If the storage time is prolonged the temperature should proprably be at
-80°.
Perhaps it's an idea to let them freeze in embedding molds. It is easy to
get the OCT-blocks out again. And then make little parafilm-parcels.

Hope this helps

Gudrun Lang
 
Biomed. Analytikerin
Histolabor
Akh Linz
Krankenhausstr. 9
4020 Linz
+43(0)732/7806-6754


-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Jamie E
Erickson
Gesendet: Donnerstag, 15. Februar 2007 23:40
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] Freezing human skin biopsies for cytokine ICC

Hello histonetters,
                                       I hope someone can help me with a 
question about how to freeze a human skin biopsy. I am in research but am 
in collaboration with a clinical lab that will be collecting human  skin 
biopsies for me to do cytokine ICC  of TNF,IFN-g, among others and 
possibly cell makers. 
       My question is how do I instruct them in freezing the skin so that 
I get the best sample for this staining. I am going to embed the sample 
once it is in my lab for cryosectioning.  Also if people know of 
manufactures of antibodies that sell  TNF,IFN-g that work in human tissue 
that would also be helpful, I have found a few but many different 
protocols and fixations.

Any thoughts how be helpful.

Jamie

_______________________________
Jamie Erickson
Sr. Research Associate 
Department: DSMP
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
508-688-3134
FAX: 508-793-4895
e-mail: jamie.erickson <@t> abbott.com
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