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Correcting the axial shrinkage of skeletal muscle thick sections visualized by confocal microscopy
Author(s) -
JANÁČEK J.,
KREFT M.,
ČEBAŠEK V.,
ERŽEN I.
Publication year - 2012
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.1365-2818.2011.03594.x
Subject(s) - microtome , confocal , deformation (meteorology) , optical microscope , microscope , stack (abstract data type) , materials science , confocal microscopy , microscopy , optical sectioning , shrinkage , skeletal muscle , anatomy , optics , biomedical engineering , composite material , scanning electron microscope , physics , biology , computer science , medicine , programming language
Summary Confocal microscopy is a suitable method for measurements and visualization of skeletal muscle fibres and the neighbouring capillaries. When using 3D images of thick sections the tissue deformation effects should be avoided. We studied the deformation in thick sections of the rat skeletal muscle from complete stacks of images captured with confocal microscope. We measured the apparent thickness of the stacks and compared it to the slice thickness deduced from calibrated microtome settings. The ratio of both values yielded the axial scaling factor for every image stack. Careful sample preparation and treatment of the tissue cryosections with cold Ringer solution minimize the tissue deformation. We conclude that rescaling by the inverse of the axial scaling factor of the stack of optical slices in the direction of the microscope optical axis satisfactorily corrects the axial deformation of skeletal muscle samples.