[Histonet] Re: Histonet Digest, Vol 110, Issue 15

[Madeleine Huey]

Colleen,
You can pre-warm your OCT block inside the cryostat @ -20c (~ hr) from
freezer before cutting, then gradually increase the temperature to
warmer with 1 degree increment (thickness dependant).  I also
pre-chilled my slides before used.

Another alternative;
- Fix with 4% PFA/PBS for ~ 1 hr on ice (you can freezed aliquot 4%PFA
@ -20c so you don't need to make them fresh every time)
- Wash 3xPBS buffer to remove excess 4% PFA/PBS, ph 7.4
- Transfer pre-fix zebra fish to 15% & 30% sucrose (30 min each or
longer, not crucial), or you can leave them in 30% sucrose & store in
fridge without freezing in OCT
- Embed with OCT on ICE, not liquid nitrogen
- Pre-warm -20c before cutting

Good Luck!
Madeleine Huey, BS, HTL & QIHC (ASCP)
Supervisor - Pathology (IPOX & Histology)
madeleine_h <@t> elcaminohospital.org


Message: 1
Date: Thu, 10 Jan 2013 15:16:32 -0600
From: Colleen Forster <cforster <@t> umn.edu>
Subject: [Histonet] zebra fish experts......
To: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <50EF2FB0.6030908 <@t> umn.edu>
Content-Type: text/plain; charset="iso-8859-1"

Anyone out there doing zebra fish work I need your help! We are trying
to fix zebra fish kidneys in 10% formalin and then freeze...the samples
just seem to crumble when we try to cut...can someone help me out here
with fixing/freezing methods for these little buggers!!

Thanks in advance!!

Colleen Forster
U of MN