Image-based analysis of living mammalian cells using label-free 3D refractive index maps reveals new organelle dynamics and dry mass flux.
Holo-tomographic microscopy (HTM) is a label-free microscopy method reporting the fine changes of a cell's refractive indices (RIs) in three dimensions at high spatial and temporal resolution. By combining HTM with epifluorescence, we demonstrate that mammalian cellular organelles such as lipid...
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| Format: | Article |
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Public Library of Science (PLoS)
2019-12-01
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| Series: | PLoS Biology |
| Online Access: | https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.3000553&type=printable |
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| author | Patrick A Sandoz Christopher Tremblay F Gisou van der Goot Mathieu Frechin |
| author_facet | Patrick A Sandoz Christopher Tremblay F Gisou van der Goot Mathieu Frechin |
| author_sort | Patrick A Sandoz |
| collection | DOAJ |
| description | Holo-tomographic microscopy (HTM) is a label-free microscopy method reporting the fine changes of a cell's refractive indices (RIs) in three dimensions at high spatial and temporal resolution. By combining HTM with epifluorescence, we demonstrate that mammalian cellular organelles such as lipid droplets (LDs) and mitochondria show specific RI 3D patterns. To go further, we developed a computer-vision strategy using FIJI, CellProfiler3 (CP3), and custom code that allows us to use the fine images obtained by HTM in quantitative approaches. We could observe the shape and dry mass dynamics of LDs, endocytic structures, and entire cells' division that have so far, to the best of our knowledge, been out of reach. We finally took advantage of the capacity of HTM to capture the motion of many organelles at the same time to report a multiorganelle spinning phenomenon and study its dynamic properties using pattern matching and homography analysis. This work demonstrates that HTM gives access to an uncharted field of biological dynamics and describes a unique set of simple computer-vision strategies that can be broadly used to quantify HTM images. |
| format | Article |
| id | doaj-art-806f1717ea574a8fbdc790d4e2896f38 |
| institution | OA Journals |
| issn | 1544-9173 1545-7885 |
| language | English |
| publishDate | 2019-12-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Biology |
| spelling | doaj-art-806f1717ea574a8fbdc790d4e2896f382025-08-20T02:18:24ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852019-12-011712e300055310.1371/journal.pbio.3000553Image-based analysis of living mammalian cells using label-free 3D refractive index maps reveals new organelle dynamics and dry mass flux.Patrick A SandozChristopher TremblayF Gisou van der GootMathieu FrechinHolo-tomographic microscopy (HTM) is a label-free microscopy method reporting the fine changes of a cell's refractive indices (RIs) in three dimensions at high spatial and temporal resolution. By combining HTM with epifluorescence, we demonstrate that mammalian cellular organelles such as lipid droplets (LDs) and mitochondria show specific RI 3D patterns. To go further, we developed a computer-vision strategy using FIJI, CellProfiler3 (CP3), and custom code that allows us to use the fine images obtained by HTM in quantitative approaches. We could observe the shape and dry mass dynamics of LDs, endocytic structures, and entire cells' division that have so far, to the best of our knowledge, been out of reach. We finally took advantage of the capacity of HTM to capture the motion of many organelles at the same time to report a multiorganelle spinning phenomenon and study its dynamic properties using pattern matching and homography analysis. This work demonstrates that HTM gives access to an uncharted field of biological dynamics and describes a unique set of simple computer-vision strategies that can be broadly used to quantify HTM images.https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.3000553&type=printable |
| spellingShingle | Patrick A Sandoz Christopher Tremblay F Gisou van der Goot Mathieu Frechin Image-based analysis of living mammalian cells using label-free 3D refractive index maps reveals new organelle dynamics and dry mass flux. PLoS Biology |
| title | Image-based analysis of living mammalian cells using label-free 3D refractive index maps reveals new organelle dynamics and dry mass flux. |
| title_full | Image-based analysis of living mammalian cells using label-free 3D refractive index maps reveals new organelle dynamics and dry mass flux. |
| title_fullStr | Image-based analysis of living mammalian cells using label-free 3D refractive index maps reveals new organelle dynamics and dry mass flux. |
| title_full_unstemmed | Image-based analysis of living mammalian cells using label-free 3D refractive index maps reveals new organelle dynamics and dry mass flux. |
| title_short | Image-based analysis of living mammalian cells using label-free 3D refractive index maps reveals new organelle dynamics and dry mass flux. |
| title_sort | image based analysis of living mammalian cells using label free 3d refractive index maps reveals new organelle dynamics and dry mass flux |
| url | https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.3000553&type=printable |
| work_keys_str_mv | AT patrickasandoz imagebasedanalysisoflivingmammaliancellsusinglabelfree3drefractiveindexmapsrevealsneworganelledynamicsanddrymassflux AT christophertremblay imagebasedanalysisoflivingmammaliancellsusinglabelfree3drefractiveindexmapsrevealsneworganelledynamicsanddrymassflux AT fgisouvandergoot imagebasedanalysisoflivingmammaliancellsusinglabelfree3drefractiveindexmapsrevealsneworganelledynamicsanddrymassflux AT mathieufrechin imagebasedanalysisoflivingmammaliancellsusinglabelfree3drefractiveindexmapsrevealsneworganelledynamicsanddrymassflux |