Re: [Histonet] Microtome calibration?

From:Rene J Buesa

Many years ago there were some methods developed to determine the thickness of sections, but changes in temperature of the wax after each section (due to friction) and the fact that wax expands with heat make those calibrations rather difficult.
  The best way is to realize the presence of layers y a section. Usually very thin sections will show an abundance of sectioned nuclei, and seldom more than one layer of lymphocytes, whose average diameter if 5-7 Ám
  If you are getting sections with well defined single layers of either RBC of lymphocytes, you are sectioning thin, if there are several layers of RBC or lymphocytes, you are sectioning on the thick side.
  Additionally, the microtome you mentioned  is quite good in delivering the thickness it is set to work.
  RenÚ J. wrote:
  Good morning,
A question was raised as to "how do you know the sections are the 
appropriate thickness?". Does one trust the settings on the microtome (Leitz 1512)? 
Is it an experienced eye that knows when they are "right"? Or is there some 
instrument that will do this measurment? Is there some way to calibrate the 
microtome to ensure that the sections actually are Xum?


Albert C. Grobe, PhD
International Heart Institute of Montana Foundation
Tissue Engineering Lab, Saint Patrick Hospital

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