[Histonet] prevent wrinkles when cutting

[Rachel M Gonzalez]

Hi

The long ribbons are for R&D and QA purposes. Often we test under multiple
conditions 1-4 with multiple antibodies to the same tissue or multiple
tissue to optimize reagents.  It quickly adds up.

Rachel


On Mon, Apr 20, 2015 at 3:28 PM, Joelle Weaver <joelleweaver <@t> hotmail.com>
wrote:

> Why are you cutting such a long ribbon? You usually only a need a series
> of 3-4 sections even for ribbon cutting. Might be easier to control if you
> don't try to move such a long ribbon to the waterbath. Drag the shorter
> ribbon towards you on the waterbath. Make sure the water is not too cool.
> Face the block to full face but superficial, chill on ice for some
> moisture, take sections while the block is still very cold. Use a slow,
> steady, smooth stroke if doing manual cutting. Make sure your embedding
> works well for the way you orient the block in the holder. Angles work well
> for many tissues that are prone to wrinkling. Its mostly just practice
> though. The more you cut, the easier it becomes and usually the better you
> get at it.
>
>
> Joelle Weaver MAOM, HTL (ASCP) QIHC
>
>
>
>
>
> > Date: Mon, 20 Apr 2015 19:06:17 +0200
> > From: j.benavides <@t> eae.csic.es
> > To: histonet <@t> lists.utsouthwestern.edu
> > Subject: Re: FW: [Histonet] prevent wrinkles when cutting
> >
> > Hi there,
> >
> > I´m curious about the soaking thing. We have never done it in our lab.
> > Which is the purpose to do it?
> >
> > Than, after facing the blocks, we chill them in a cold plate so, if
> > wanting to do the soaking , when should we? I guess before placing them
> > on the cold plate, but that may cause a bit of ice formation?
> >
> > Thanks a lot for your help
> >
> > Julio
> >
> >
> > On 20/04/2015 18:40, Grantham, Andrea L - (algranth) wrote:
> > > Rachel,
> > > First off, are you chilling and soaking the blocks after you face them?
> > > Do that and see if there is a difference.
> > > Don't try to get many sections to your ribbons. Shoot for a smaller
> ribbon (5-6) sections that are good. Cut slowly but consistently.
> > > What microtome are you using? Are you using disposable blades and are
> they sharp? Don't expect them to cut well if you use the same blade to face
> the blocks. If you aren't using disposables, get some! They will make your
> life easier.
> > > You might try to find a histotech at a local hospital lab who might be
> able to give you a hands-on lesson.
> > > Don't despair! We all sat down at our microtomes those first times and
> suffered trying to get perfect sections. It takes practice. You might make
> some blank blocks or blocks with tissue you can spare to practice your
> cutting techniques. I used to do this with my students and it really helped
> them.
> > > Good luck!
> > >
> > > Andi G.
> > > ________________________________________
> > > From: histonet-bounces <@t> lists.utsouthwestern.edu [
> histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of Rachel M Gonzalez
> [rachel <@t> gbi-inc.com]
> > > Sent: Monday, April 20, 2015 9:07 AM
> > > To: histonet <@t> lists.utsouthwestern.edu
> > > Subject: [Histonet] prevent wrinkles when cutting
> > >
> > > Hi
> > >
> > > Thursday was the first time I ever used a microtome.... I move to a lab
> > > that does not have someone dedicated to cutting. I already miss her.
> > >
> > > I have no problems getting ribbons of 10-30 sections long but the
> pieces
> > > are half the size of the original block. I am guessing they are
> wrinkling.
> > > What am I doing wrong?
> > >
> > > Thanks
> > > Rachel
> > > Senior Scientist....
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