Applying the Expansion Microscopy Method in Neurobiology
2021
Many biological studies require the analysis of ultrastructural
changes at the level of cell organelles and macromolecules. Since
the resolution of modern confocal microscope is limited by the diffraction
limit (200–300 nm), it is impossible to study such small objects
using standard fluorescence microscopy. Ultra-high resolution microscopy
methods require expensive equipment and are technically difficult
in use, which in turn limits their widespread practical application. However,
recently appeared methods make it possible to increase the resolution
of microscopy not by improving the image registration system, but
by physically isotropic expansion of a biological sample using a
controlled chemical process. Due to this method, called expansion
or expansive microscopy (ExM), it became possible to obtain three-dimensional
images of samples with a resolution sufficient to study individual
cell organelles using a conventional confocal microscope. This review
covers the history of this method, its basic principles and examples
of use in various fields of biology and medicine, as well as reflects
future directions for improving this technology. The article discusses
the methodological features of the ExM application in a study of
brain tissue samples using the algorithm that allows adaptation
of the standard protocol to the goals and objectives of a particular
study.
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