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Organelle-specific phase contrast microscopy enables gentle monitoring and analysis of mitochondrial network dynamics
Author(s) -
Siyue Guo,
Ying Ma,
Yang Pan,
Zachary J. Smith,
Kaiqin Chu
Publication year - 2021
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.425848
Subject(s) - organelle , mitochondrion , photobleaching , fluorescence recovery after photobleaching , fluorescence microscope , microscopy , live cell imaging , biophysics , microbiology and biotechnology , biological system , fluorescence , chemistry , biology , cell , biochemistry , physics , optics
Mitochondria are delicate organelles that play a key role in cell fate. Current research methods rely on fluorescence labeling that introduces stress due to photobleaching and phototoxicity. Here we propose a new, gentle method to study mitochondrial dynamics, where organelle-specific three-dimensional information is obtained in a label-free manner at high resolution, high specificity, and without detrimental effects associated with staining. A mitochondria cleavage experiment demonstrates that not only do the label-free mitochondria-specific images have the required resolution and precision, but also fairly include all cells and mitochondria in downstream morphological analysis, while fluorescence images omit dim cells and mitochondria. The robustness of the method was tested on samples of different cell lines and on data collected from multiple systems. Thus, we have demonstrated that our method is an attractive alternative to study mitochondrial dynamics, connecting behavior and function in a simpler and more robust way than traditional fluorescence imaging.

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