[Histonet] Thick cryosections

[Patsy Ruegg]

Yes this was another method I was going to suggest, rather tedious isn't it?


I have also had some success using elmers glue coated slides for things like
this, I make them myself, 5% glue in dih20, dip the non charged slides in
the glue then airdry, store them dry, then when you use them as they dip in
water it seems to reactivate the glue and things stick, I have used this
with success with GMA embedded cartilage and bone, cartilage especially
tends to peel off the slide, there are so few cells and no vessels to attach
to the slide with cartilage so it is a problem.  It also helps to dry the
slides flat on a slide warmer at about 40dc overnight, higher temps make the
cartilage dislodge. 

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net 
www.ihcrg.org


-----Original Message-----
From: Montina Van Meter [mailto:Montina.VanMeter <@t> pbrc.edu] 
Sent: Thursday, October 13, 2011 9:46 AM
To: Patsy Ruegg; Carmen Maria Garcia Pascual;
histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Thick cryosections

Carmen,

I routinely stain 40-60 micron sections for IHC.  I currently run all my
IHC with a free-floating protocol which allows for maximum staining
penetration from both sides of the section and then mount them on the
slides.  In the past, we mounted the 60um sections on (300 bloom)
gelatin coated slides that were processed by hand in a humidity chamber.
If you would like additional information about this technique, please
send me an email.

Regards,

Tina 







Montina J. Van Meter, HT (ASCP)
Lab Manager
Dept. of Autonomic Neuroscience
Pennington Biomedical Research Center
6400 Perkins Rd.
Baton Rouge, LA  70791
225-763-2564


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Patsy
Ruegg
Sent: Thursday, October 13, 2011 9:30 AM
To: 'Carmen Maria Garcia Pascual'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Thick cryosections

I don't care what kink of slides you use if you cut 50 micron thick
sections and try to do IHC you are going to lose some or all sections,
in my experience.  The only thing that might work and even this is
questionable because of the thickness is the tape transfer system but I
really doubt you are going to be able to get sections that thick to
stick to the polymer coated slide and not come off with the tape.

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net
www.ihcrg.org


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Carmen
Maria
Garcia Pascual
Sent: Tuesday, October 11, 2011 9:11 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Thick cryosections


Good afternoon everyone!

I have made 50um mice ovarian cryosections and I have used superfrost 
plus slides. But some of them fall of the slides when I perfomed the 
immuno, one colleague has told me that this is because the cuts are too 
thick, and I think she is rigth. I don't know if there is some thing 
that I can make to avoid this, like using another kind of slides...

Thanks!!!!!!!!!!!


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