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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| http://dx.doi.org/10.1111/boc.202400038 | DOI Listing |
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