Cryoconveyor Protocols in Correlative Light and Electron Microscopy: From Multilevel Imaging to Modeling the Biophysical Effects and “Cryotheranostics”
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Abstract
This paper is a technical and methodological note aimed to introduce into the practice of biological research methods a cryomicroscopy in a conveyor mode from small magnifications to the limits of magnification/resolution of scanning electron cryomicroscopy (CryoSEM). The protocol described can be applied to the samples of a low sample preparation complexity without ultratomy or processing typical of transmission electron microscopy. According to this protocol, samples are analyzed in a single microcuvette (chip) indexed by laboratory information management system and sequentially transferred from the nondestructive low-resolution optical microscopy instruments, such as lensless cryo-microscopes, to the CryoSEM/CryoESEM level (in programmable environments and atmospheres). Methods that were introduced and tested include correlative lensless cryomicroscopy and CryoSEM (including those with the sequential transition to microanalysis on wavelength-dispersive X-ray spectrometer on the Rowland circle) as well as microscopy and microinterferometry methods in the ranges from infrared to far-ultraviolet. Among the advantages of the cryoconveyor analysis protocols are sample preservation in a single portable cuvette-chip along with a possibility to establish spatial colocalization between data of optical and electron microscopy using pattern recognition software (all the way to indexing in the laboratory information system) in conducting a full range of microscopic examination. An opportunity is additionally provided for a comprehensive nondestructive sample analysis in a sequential study of microscopic systems with a possibility of variation at the subsequent stages of high-resolution microscopy, depending on the results obtained at the preceding stages of microscopy (at lower resolution).
期刊介绍：
Moscow University Biological Sciences Bulletin is forum for research in all important areas of modern biology. It publishes original work on qualitative, analytical and experimental aspects of research. The scope of articles to be considered includes plant biology, zoology, ecology, evolutionary biology, biophysics, genetics, genomics, proteomics, molecular biology, cell biology, biochemistry, endocrinology, immunology, physiology, pharmacology, neuroscience, gerontology, developmental biology, bioinformatics, bioengineering, virology, and microbiology.
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