What is it about?
Under certain circumstances, cells could remain uncut in cryostat slices, protruding over the free cut surface. Such phenomenon may be assessed by means of confocal microscopy.
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Why is it important?
Those cell protrusions also affect the estimation of section thickness, which is later needed for the application of stereological methods. If section thickness was overestimated, the calculations of cell populations based on that thickness would be overestimated too.
Perspectives
If precision in cell counts is desired, researchers are encouraged to check if the phenomenon occurs in their material by confocal microscopy and to apply the formula proposed in the article to calculate the degree of the overestimation (Maximal thickness/ geometrical thickness). They are also to perform the calculations the same day of cutting since progressive reduction of tissue thickness of cryostat slices occurs if stored in no-frost refrigerators or freezers (up to 50% after 30 days).
Anna Puigdellivol
Universitat de Barcelona
Read the Original
This page is a summary of: Cryostat Slice Irregularities May Introduce Bias in Tissue Thickness Estimation: Relevance for Cell Counting Methods, Microscopy and Microanalysis, July 2015, Cambridge University Press,
DOI: 10.1017/s143192761501380x.
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