[Histonet] prevent wrinkles when cutting

[Pam Marcum]

The one question we have not asked is; are you cutting serial sections for these blocks?  If you are are you laying them out on a board and not a waterbath?  I have done that with neuropath sections in research for whole rat and monkey brains.  Then your ribbon length would make sense.  Generally the longer the ribbon you have to handle the more likely you are to lose sections or have it fold over on its self.  When you are just learning mastering shorter ribbons is easier.  If you are going to a waterbath you can crowd the ribbon on the bath and it had no place to gently stretch out. 
  
Pam Marcum 

----- Original Message -----

From: "Joelle Weaver" <joelleweaver <@t> hotmail.com> 
To: "Julio Benavides" <j.benavides <@t> eae.csic.es>, "Histonet" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Monday, April 20, 2015 2:28:47 PM 
Subject: RE: [Histonet] prevent wrinkles when cutting 

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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