Can you trust your cryostat? Reproducibility of cryostat section thickness.
Reproducibility of cryostat section thickness is required for valid quantitative microscopy. This is generally pursued by motorized sectioning using a low but constant speed. The purpose of our study was to compare variation in section thickness between motorized and manual cryostat sectioning. Serial sections were cut from a frozen block of homogenized tissue on different days. Lactate dehydrogenase activity was histochemically detected and calibrated absorbance measurements were taken. The coefficients of variation of measurements was 9.7% for motorized sectioning and 3.3% for manual sectioning. In conclusion, section thickness is similarly reproducible after manual sectioning compared with motorized sectioning, if not better.
De Witt Hamer
Philip C
PC
Department of Neurosurgery, Academic Medical Center, University of Amsterdam, The Netherlands. P.C.deWittHamer@amc.nl
Bleeker
Fonnet E
FE
Zwinderman
Aeilko H
AH
Van Noorden
Cornelis J F
CJ
D000818:Animals; D001331:Automation / Q000592:standards; D016616:Cryoultramicrotomy / Q000592:standards*; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008099:Liver / Q000166:cytology; D051381:Rats; D015203:Reproducibility of Results
D007770:L-Lactate Dehydrogenase