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Tomographic phase microscopy

Nature Methods volume 4, pages 717–719 (2007)Cite this article

Abstract

We report a technique for quantitative three-dimensional (3D) mapping of refractive index in live cells and tissues using a phase-shifting laser interferometric microscope with variable illumination angle. We demonstrate tomographic imaging of cells and multicellular organisms, and time-dependent changes in cell structure. Our results will permit quantitative characterization of specimen-induced aberrations in high-resolution microscopy and have multiple applications in tissue light scattering.

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Figure 1: Tomographic phase microscope.
Figure 2: Refractive index tomogram of a HeLa cell.
Figure 3: Effects of acetic acid on a HeLa cell and imaging of C. elegans.

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Acknowledgements

This work was funded by US National Institutes of Health (P41-RR02594-18) and Hamamatsu Corporation.

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Authors and Affiliations

  1. G.R. Harrison Spectroscopy Laboratory, 77 Massachusetts Avenue 6-014, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Wonshik Choi, Christopher Fang-Yen, Kamran Badizadegan, Seungeun Oh, Niyom Lue, Ramachandra R Dasari & Michael S Feld

  2. Department of Pathology, Harvard Medical School and Massachusetts General Hospital, Boston, 02114, Massachusetts, USA

    Kamran Badizadegan

Authors
  1. Wonshik Choi
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  2. Christopher Fang-Yen
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  3. Kamran Badizadegan
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  4. Seungeun Oh
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  5. Niyom Lue
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  6. Ramachandra R Dasari
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  7. Michael S Feld
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Contributions

W.C. and C.F.-Y. conducted the experimental setup, data acquisition and data analysis; S.O. contributed to the experimental setup; N.L. contributed to 3D visualization; W.C., C.F.-Y., K.B., R.R.D. and M.S.F. wrote the manuscript; M.S.F. supervised the project.

Corresponding author

Correspondence to Michael S Feld.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Methods, Supplementary Software (PDF 911 kb)

Supplementary Video 1

Animation of 3-D rendered index tomogram of a HeLa cell from Figure 2. (MOV 2460 kb)

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Choi, W., Fang-Yen, C., Badizadegan, K. et al. Tomographic phase microscopy. Nat Methods 4, 717–719 (2007). https://doi.org/10.1038/nmeth1078

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