Image-based analysis of living mammalian cells using label-free 3D refractive index maps reveals new organelle dynamics and dry mass flux.

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  • Additional Information
    • Source:
      Publisher: Public Library of Science Country of Publication: United States NLM ID: 101183755 Publication Model: eCollection Cited Medium: Internet ISSN: 1545-7885 (Electronic) Linking ISSN: 15449173 NLM ISO Abbreviation: PLoS Biol Subsets: MEDLINE
    • Publication Information:
      Original Publication: San Francisco, CA : Public Library of Science, [2003]-
    • Subject Terms:
      Microscopy, Fluorescence/*methods ; Organelles/*physiology ; Refractometry/*methods; HeLa Cells ; Humans ; Image Processing, Computer-Assisted/methods ; Imaging, Three-Dimensional/methods ; Lipid Metabolism ; Mitochondria/metabolism
    • Abstract:
      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.
      Competing Interests: I have read the journal's policy, and the authors of this manuscript have the following competing interests: Dr. Mathieu Fréchin is an employee of Nanolive SA.
    • References:
      Sci Rep. 2016 Nov 22;6:36815. (PMID: 27874018)
      Appl Opt. 1999 Dec 1;38(34):6994-7001. (PMID: 18324243)
      Science. 1986 Nov 14;234(4778):863-6. (PMID: 3775367)
      Elife. 2019 May 07;8:. (PMID: 31063131)
      J Cell Sci. 2008 Oct 1;121(Pt 19):3187-95. (PMID: 18782860)
      Exp Cell Res. 1986 Apr;163(2):525-38. (PMID: 3514249)
      J Cell Biol. 2013 Dec 23;203(6):985-1001. (PMID: 24368806)
      Dev Cell. 2013 Feb 25;24(4):384-99. (PMID: 23415954)
      Hum Mol Genet. 2018 Jun 15;27(12):2101-2112. (PMID: 29659804)
      Nat Cell Biol. 1999 Jun;1(2):113-8. (PMID: 10559883)
      J Biomed Opt. 2006 Sep-Oct;11(5):054032. (PMID: 17092181)
      Opt Express. 2010 Sep 13;18(19):19462-78. (PMID: 20940842)
      Nat Methods. 2007 Sep;4(9):717-9. (PMID: 17694065)
      Exp Cell Res. 1988 Apr;175(2):409-13. (PMID: 3360061)
      Cardiovasc Diabetol. 2005 Jan 10;4:2. (PMID: 15642122)
      Appl Opt. 2018 Jul 1;57(19):5364-5370. (PMID: 30117828)
      J Biol Chem. 2003 Sep 19;278(38):36373-9. (PMID: 12783892)
      Cell Adh Migr. 2014;8(1):42-8. (PMID: 24589621)
      Elife. 2014;3:e01607. (PMID: 24497546)
      J Opt Soc Am A Opt Image Sci Vis. 2007 Jun;24(6):1617-22. (PMID: 17491629)
      PLoS One. 2008 Jan 23;3(1):e1477. (PMID: 18213385)
      Anal Sci. 2016;32(2):125-7. (PMID: 26860553)
      Exp Cell Res. 1953 Sep;5(1):191-6. (PMID: 13083628)
      J Biophotonics. 2016 Oct;9(10):1068-1076. (PMID: 27010098)
      Phys Rev Lett. 2012 Sep 14;109(11):118105. (PMID: 23005682)
      J Cell Biol. 2015 Nov 23;211(4):757-63. (PMID: 26598613)
      Mol Metab. 2015 Dec 31;5(3):153-163. (PMID: 26977387)
      Nat Commun. 2019 Jan 21;10(1):340. (PMID: 30664642)
      Cell Cycle. 2007 Jul 1;6(13):1613-20. (PMID: 17581282)
      Curr Opin Cell Biol. 2014 Aug;29:39-45. (PMID: 24736091)
      Curr Opin Genet Dev. 2011 Oct;21(5):585-90. (PMID: 21945002)
      J Cell Biol. 2015 Nov 23;211(4):765-74. (PMID: 26598614)
      Lipids. 2009 Jun;44(6):477-87. (PMID: 19440746)
      Traffic. 2012 May;13(5):705-14. (PMID: 22300234)
      Methods Cell Biol. 2008;90:xvii-xix. (PMID: 19195542)
      Nat Methods. 2018 Oct;15(10):751. (PMID: 30275585)
      Nature. 2011 Jan 13;469(7329):226-30. (PMID: 21196934)
      Opt Express. 2009 Jul 20;17(15):13080-94. (PMID: 19654713)
      Semin Cell Dev Biol. 2018 Oct;82:41-50. (PMID: 29241691)
      J Lipid Res. 2013 Dec;54(12):3419-29. (PMID: 24103784)
      Ultrason Imaging. 1982 Oct;4(4):336-50. (PMID: 6891131)
      Sci Adv. 2019 Jun 26;5(6):eaav9960. (PMID: 31249869)
      Nat Methods. 2014 Dec;11(12):1221-8. (PMID: 25423019)
      Nat Methods. 2012 Jul;9(7):671-5. (PMID: 22930834)
      PLoS One. 2015 Mar 10;10(3):e0121645. (PMID: 25756421)
      Z Zellforsch Mikrosk Anat. 1955;41(5):481-92. (PMID: 14387135)
      Genome Biol. 2006;7(10):R100. (PMID: 17076895)
      Curr Opin Cell Biol. 2017 Aug;47:9-15. (PMID: 28231490)
    • Publication Date:
      Date Created: 20191220 Date Completed: 20200320 Latest Revision: 20200320
    • Publication Date:
      20221213
    • Accession Number:
      PMC6922317
    • Accession Number:
      10.1371/journal.pbio.3000553
    • Accession Number:
      31856161