Affiliation:
1. N.N. Semenov Federal Research Center for Chemical Physics, Russian Academy of Sciences, Institute of Biochemical Physics, Russian Academy of Sciences
Abstract
This paper is a technical and methodological note, the purpose of which is to introduce into
the practice of biological research methods of cryomicroscopy in a conveyor mode, starting
from small magnifications and ending with the limits of magnification/resolution of scanning
electron cryomicroscopy. The protocol described can be applied to the samples with low
sample preparation complexity without ultratomy or the sample processing typical for transmission electron microscopy methods. According to this protocol samples are analyzed in
a single microcuvette (chip) indexed by laboratory information management system and
sequentially moved from the non-destructive low-resolution optical microscopy instruments
(such as lensless cryomicroscopes) and optical super-resolution methods (some
microinterferometers and MIMs with cryotables) to the CryoSEM/CryoESEM level
(in programmable environments and atmospheres). Methods of correlation lensless
cryomicroscopy and scanning microscopy (including those with the subsequent transition to
microanalysis) were introduced; CryoCUVEM and CryoCIREM methods in the ultraviolet and
infrared range, respectively; microinterferometry methods using a multi-beam reflected light
interferometer (based on the MII-11 platform with several changes); the development of
CryoCDICEM systems based on the optical path of an inverted metallographic microscope
with a DIC attachment and a LED emitter was also initiated. The advantages of cryoconveyor
analysis protocols are ensuring the sample safety in a single cuvette-chip and the possibility of
establishing spatial colocalization between the data of optical and electron microscopy
(including in the CLEM/CryoCLEM mode), as well as providing a comprehensive non-
destructive sample analysis in the sequential study of the microscopic systems with the
possibility of varying the subsequent stages of high-resolution microscopy, depending on the
results obtained at the previous stages of lower resolution microscopic studies.
Cited by
1 articles.
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