[Histonet] Reference to microtome micrometer thickness

[Elizabeth Chlipala]

Reuel

When your microtomes are serviced yearly they should be calibrated at the same time for micron thickness.   We have Leica microtomes and they are serviced by one of their vendors.  I had actually asked our service technician if they check for that and he said that do that, he also pointed out that your knife angle can also affect section thickness.  But that is just calibrating the microtome.  Overall section thickness can vary dependent upon many variables.

The type of disposable blade you use, how sharp that blade is, how many blocks you have sectioned with that particular blade,  how cold your block is, how fast or slow you turn the wheel of your microtome, some people blow on the blocks, which we all know makes sectioning easier, because it warms up the block and therefore you will get a thicker section, all of those parameters can affect your micron thickness.  Slight variation in section thickness may not impact routine H&E slides for diagnosis, etc.  But when you are running IHC, or any stain and then utilizing image analysis that's where section thickness consistency is very important.

If a section is too thick you will be able to focus through multiple planes of nuclei in the sample, meaning you should see only a  single cell layer when tissue has been cut at the proper thickness.  

I hope this helps.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
liz <@t> premierlab.com
www.premierlab.com

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Reuel Cornelia
Sent: Friday, November 21, 2014 4:20 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Reference to microtome micrometer thickness

Hello Histonetters,
how will we verify the thickness of a paraffin section in a slide. Do we have a reference regarding  on how to measure the thickness. I based the  thickness of my section thru the mcirometer on the rotary microtome but one of our reveiwers does not believed that we are cutting them into
3 micron thickness or 4 micron thickness. Please if we have any reference, please share it to me. 
 
 
 
 
Reuel Cornelia, BS MT, AMT
Cellular Pathology
Texas Scottish Rite Hospital for Children
2222 Welborn Street
Dallas, TX 75219
Tel: 214-559-7766
fax: 214-559-7768