[Histonet] paraffin sectioning-dry tissue?

[Lucie Guernsey]

By adding water, ice, or warm humidity (through exhalations) to the mix,
though, doesn't the block contract/expand? Wouldn't that change the
ultimate thickness of the section? I've always wondered how much it affects
things.

Could your sections be 1 um (or multiple um!) thicker/thinner that you
expect? What happens if individual blocks contract/expand differently (due
to the amount of time left soaking, how far into the block you've cut since
you last soaked, etc.)? I feel like you couldn't properly compare
quantifications across a study if the thickness of your tissue is an
unknown variable.

Am I overthinking this?

Lucie Guernsey
UC San Diego
(858) 822-5797
lguernsey <@t> ucsd.edu


On Thu, Feb 5, 2015 at 7:35 AM, James Watson <JWatson <@t> gnf.org> wrote:

> We use a 5% glycerin in denatured alcohol for our 100% alcohol on our
> tissue processor for routine animal tissues, this reduces the over
> dehydration of the animal tissue and greatly reduces the time required to
> soak the blocks.  Warning, if processing fat or cell pellets do not use
> this, we switch the containers to straight 100% reagent alcohol for them.
> For fat we have a longer processing schedule and for cell pellets we have a
> short processing cycle.
>
> James Watson HT  ASCP
> GNF  Genomics Institute of the Novartis Research Foundation
> Scientific Technical Leader II, Histology
> Tel    858-332-4647
> Fax   858-812-1915
> jwatson <@t> gnf.org
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:
> histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Caroline Miller
> Sent: Thursday, February 05, 2015 6:41 AM
> To: Geoff
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] paraffin sectioning-dry tissue?
>
> Yes, exactly what Mike and Geoff said. All mouse tissue, especially liver,
> can be really dry and needs a 'soak'. I have left them for an hour before
> now but don't leave it for longer than 4 hours though because it can start
> to swell and de-process!
>
> You will still only get a few non-chattery sections so be gentle. Thinner
> sections also help too (3-4.5). Plus low um polishing after you trim
>
> Good luck! It is weird at first but you will get used to it!
>
> Caroline
>
> Sent from my iPhone
>
> > On Feb 5, 2015, at 6:29 AM, Geoff <mcauliff <@t> rwjms.rutgers.edu> wrote:
> >
> > This is common with mouse and rat tissues, they get "over-dried" with a
> typical processing schedule.
> > Soaking the face of the block with a kimwipe wet with ice water for 60
> -120 seconds will enable you to cut 10 nice sections, maybe more.
> >
> > Geoff
> >
> >> On 2/5/2015 6:23 AM, Emily Brown wrote:
> >> Hello all!
> >>
> >> I just started sectioning mouse liver in paraffin and the tissue is
> >> very dry.  I know it's not supposed to have water due to the
> >> processing, but the weird thing is that one tech's solution is to put
> >> a wet kimwipe on the block for a while.
> >> It seems to me that there is a larger processing issue if this is
> >> happening, am I correct? And why add water when you've already
> >> dehydrated it?
> >> Unfortunately, we do not have the set up to embed them ourselves, so
> >> we have to send them to a histology lab.  They were sectioning for
> >> us, but they are backlogged, so my boss wants me to do it.
> >> Therefore, I can't tell you how they were processed, but I think
> >> usually the histology lab manages to get good sections.
> >> Is putting a wet kimwipe (using distilled water) the best way to get
> >> rid of chatter that's only in the tissue? The surrounding paraffin
> >> sections excellent.
> >> This may have been answered already, but a very quick google search
> >> didn't help.  My googlefu is probably erratic as it's still early.
> >>
> >> Emily
> >>
> >>
> >> "By bitching and bitching and bitching, they could exhaust the drama
> >> of their own horror stories. Grow bored. Only then could they accept
> >> a new story for their lives. Move forward."
> >>
> >> -Chuck Palahniuk, "Haunted"
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> >
> >
> > --
> > --
> > **********************************************
> > Geoff McAuliffe, Ph.D.
> > Neuroscience and Cell Biology
> > Robert Wood Johnson Medical School
> > 675 Hoes Lane, Piscataway, NJ 08854
> > voice: (732) 235-4583; fax: -4029
> > mcauliff <@t> rwjms.rutgers.edu
> > **********************************************
> >
> >
> >
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