Advancing yeast cell analysis: A cryomethod for serial block-face scanning electron microscopy imaging in mitochondrial morphology studies.

IF 2.4 4区 生物学 Q4 CELL BIOLOGY
Corinne Blancard, Fanny Decoeur, Stéphane Duvezin-Caubet, Marie-France Giraud, Bénédicte Salin
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引用次数: 0

Abstract

Background information: Conventional Transmission Electron Microscopy analysis of biological samples often provides limited insights due to its inherent two-dimensional (2D) nature. This limitation hampers a comprehensive understanding of the three-dimensional (3D) complexity of cellular structures, occasionally leading to misinterpretations. Serial block-face scanning electron microscopy emerges as a powerful method for acquiring high-resolution 3D images of cellular volumes. By iteratively removing ultrathin sample sections and capturing images of each newly exposed surface, Serial block-face scanning electron microscopy allows for the meticulous reconstruction of a comprehensive 3D volume.

Results: In this study, we investigate the 3D architecture of altered mitochondrial morphologies in Saccharomyces cerevisiae using Serial block-face scanning electron microscopy imaging. We have developed a novel cryomethod based on plunge freezing and a dedicated freeze-substitution protocol.

Conclusion: This protocol enhances ultrastructural preservation enabling a more accurate understanding of mitochondrial defects observed in 2D electron microscopy.

Significance: Our findings underscore the utility of Serial block-face scanning electron microscopy coupled with optimized sample preparation techniques in elucidating complex cellular structures in 3D.

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来源期刊
Biology of the Cell
Biology of the Cell 生物-细胞生物学
CiteScore
5.30
自引率
0.00%
发文量
53
审稿时长
>12 weeks
期刊介绍: The journal publishes original research articles and reviews on all aspects of cellular, molecular and structural biology, developmental biology, cell physiology and evolution. It will publish articles or reviews contributing to the understanding of the elementary biochemical and biophysical principles of live matter organization from the molecular, cellular and tissues scales and organisms. This includes contributions directed towards understanding biochemical and biophysical mechanisms, structure-function relationships with respect to basic cell and tissue functions, development, development/evolution relationship, morphogenesis, stem cell biology, cell biology of disease, plant cell biology, as well as contributions directed toward understanding integrated processes at the organelles, cell and tissue levels. Contributions using approaches such as high resolution imaging, live imaging, quantitative cell biology and integrated biology; as well as those using innovative genetic and epigenetic technologies, ex-vivo tissue engineering, cellular, tissue and integrated functional analysis, and quantitative biology and modeling to demonstrate original biological principles are encouraged.
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