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Cryofracture of paraffin‐embedded heart muscle cells
Author(s) -
Dalen Helge,
Myklebust Reidar,
Saetersdal Thv. Selmer
Publication year - 1978
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1978.tb01161.x
Subject(s) - transmission electron microscopy , liquid nitrogen , scanning electron microscope , xylene , electron microscope , materials science , endoplasmic reticulum , optical microscope , microscopy , composite material , anatomy , chemistry , biomedical engineering , pathology , nanotechnology , optics , biology , medicine , biochemistry , physics , organic chemistry , benzene
SUMMARY A comparative study of internal cellular structures of the sheep ventricular myocardium has been conducted by scanning electron microscopy (SEM) and by transmission electron microscopy (TEM). Access to the cell interior for three‐dimensional studies was obtained by cryo‐fracturing paraffin‐embedded tissue frozen in liquid nitrogen. For accurate localization of structures of special interest thick paraffin sections were examined in the light microscope (LM). Based on the information gained, it was possible to fracture the block in a desired plane. The fracturing was carried out by a light blow to a precooled scalpel held against the surface of the block, which was immersed in liquid nitrogen. After thawing and deparaffinizing at room temperature in several baths of xylene, the tissue pieces were critical point dried using CO 2 . As xylene was found to be miscible with CO 2 , it also served as an intermediate fluid. This method resulted in good preservation of the myofibrils, mitochondria, sarcoplasmic reticulum and transverse tubules (T‐tubules), which was confirmed by TEM studies of conventionally prepared tissue and of tissue originally prepared for SEM.