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Three-dimensional correlative single-cell imaging utilizing fluorescence and refractive index tomography.

Mirjam Schürmann1, Gheorghe Cojoc1, Salvatore Girardo1

  • 1Biotechnology Center of the TU Dresden, Dresden, Germany.

Journal of Biophotonics
|August 12, 2017
PubMed
Summary

Optical diffraction tomography (ODT) measures cell refractive index (RI) without markers. New optofluidic rotation (RAFTOR) imaging enables 3D RI and fluorescence correlation for cell biophysics research.

Keywords:
cell nucleusflow cytometryoptical diffraction tomographyoptical trappingrefractive indexretina cellssingle-cell analysistomography

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Area of Science:

  • Biophysics
  • Optical Imaging
  • Cell Biology

Background:

  • Cells refract light, causing aberrations in imaging.
  • Optical diffraction tomography (ODT) quantifies refractive index (RI), a key biophysical property.
  • RI measurement complements fluorescence imaging and avoids cell labeling.

Purpose of the Study:

  • Introduce a novel technique combining RI and fluorescence tomography with optofluidic rotation (RAFTOR).
  • Enable 3D segmentation and quantitative analysis of correlated RI and fluorescence data.
  • Investigate nuclear RI in suspended cells.

Main Methods:

  • Developed RAFTOR for simultaneous RI and fluorescence tomography of suspended cells.
  • Utilized optofluidic rotation for 3D data acquisition.
  • Validated the technique with cell phantoms and biological samples.

Main Results:

  • Demonstrated successful 3D segmentation of correlated RI and fluorescence data.
  • Confirmed a lower nuclear RI in HL60 cells compared to previous findings.
  • Observed nuclear inversion in adult mouse photoreceptor cells' RI distribution.

Conclusions:

  • RAFTOR provides a powerful tool for quantitative analysis of cell and tissue biophysical properties.
  • The technique advances cell imaging by correlating RI and fluorescence.
  • Findings support previous predictions and highlight RAFTOR's potential for biological discovery.