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Can you trust your cryostat? Reproducibility of cryostat section thickness
Author(s) -
De Witt Hamer Philip C.,
Bleeker Fonnet E.,
Zwinderman Aeilko H.,
Van Noorden Cornelis J.F.
Publication year - 2006
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20354
Subject(s) - cryostat , reproducibility , materials science , optical sectioning , biomedical engineering , microscopy , chemistry , chromatography , optics , medicine , physics , superconductivity , quantum mechanics
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. Microsc. Res. Tech., 2006. © 2006 Wiley‐Liss, Inc.