[Histonet] Re: cryosectioning of the lung

[Ing. Barbara Bublava]

Hi Dan,
here are the collected answers

Curtis D. King:
I soak the tung tissue in OCT and sailine 1:1 if at 4-8C overnight. This
helps to fill the spaces in the lung tissue.

Richard  Edwards
Inflate lung  with   proprietary embedding  compound.....

Vinnie Della Speranza
The next issue of HistoLogic will include a comprehensive article by Gayle
Callis on the handling of tissues intended for cryosectioning. Gayle may
respond to this query herself but have you considered perfusing the lung
with OCT before freezing? The OCT can, in this manner, occupy the spaces
filled with air that will provide the support the tissue needs during
sectioning.

Joost Bruijntjes
Do you have the opportunity to inflate this lung tissue with OCT? If not, it
will really help you.

Douglas Gregg DVM PhD
Try this, it works for me. The problem is no OCT in the alveolae. Put a
drop of OCT on the face of the block and quickly press the face against
the freezing metal block in the bottom of the crystat. This will force
some OCT into the tissue and you should be able to get at least a few
sections beofre having to repeat this procedure. Let me know if it works
for you.

and one answer, sorry - can´t find the mail right now suggested to soak
overnight in 20% sucrose.

I found a lot of answers in the archive about the cryoprotecting role of
sucrose. I did not know, that it makes the bubbles go away.

Thanks for the help. My lung sections are now real pretty ones :o)

Barbara

----- Original Message -----
From: "Dan Luchtel" <dluchtel <@t> u.washington.edu>
To: <barbara.bublava <@t> meduniwien.ac.at>
Sent: Thursday, March 18, 2004 2:05 AM
Subject: cryosectioning of the lung


> Would it be possible for you to send to me (or to the list) an e-mail
> summary of the responses as apparently, the responses were sent
> directly to you and not to the listserv?
>
> Many thanks, Dan Luchtel
>
>
>
> Message: 13
> Date: Fri, 12 Mar 2004 08:39:17 +0100
> From: "Ing. Barbara Bublava" <barbara.bublava <@t> meduniwien.ac.at>
> Subject: [Histonet] Thanks for Replys to "cryosectioning of the lung"
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <001701c40805$20182b10$9865fea9 <@t> GERICHTS9XOZZ8>
> Content-Type: text/plain; charset="iso-8859-1"
>
> I wish to thank all people who responded to my mail.
> The suggested procedures (OTC 1:1, saccarose) did work fine - at least I
> decided to go on with the saccarose for the fixed lung and a drop of OTC
on
> the surface for not fixed lung.
>
> Barbara Bublava
>
>
> --
>