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Digital holographic microtomography for high-resolution refractive index mapping of live cells

  1. Jing-Wei Su1,†,
  2. Wei-Chen Hsu1,†,
  3. Cheng-Ying Chou3,
  4. Chen-Hao Chang3,
  5. Kung-Bin Sung1,2,4,*

Article first published online: 24 AUG 2012

DOI: 10.1002/jbio.201200022

Issue

Journal of Biophotonics

Journal of Biophotonics

Volume 6, Issue 5, pages 416–424, May 2013

Additional Information

How to Cite

Su, J.-W., Hsu, W.-C., Chou, C.-Y., Chang, C.-H. and Sung, K.-B. (2013), Digital holographic microtomography for high-resolution refractive index mapping of live cells. J. Biophoton., 6: 416–424. doi: 10.1002/jbio.201200022

Author Information

  1. 1

    Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei, Taiwan, ROC

  2. 2

    Department of Electrical Engineering, National Taiwan University, Taipei, Taiwan, ROC

  3. 3

    Department of Bio-Industrial Mechatronics Engineering, National Taiwan University, Taipei, Taiwan, ROC

  4. 4

    Molecular Imaging Center, National Taiwan University, Taipei, Taiwan, ROC

  1. †

    Equal contribution.

*Phone: +886 2 3366 9675

  1. †

    Equal contribution.

Publication History

  1. Issue published online: 2 MAY 2013
  2. Article first published online: 24 AUG 2012
  3. Manuscript Accepted: 23 JUL 2012
  4. Manuscript Revised: 9 MAY 2012
  5. Manuscript Received: 13 FEB 2012

Funded by

  • National Science Council in Taiwan. Grant Number: NSC 98-2112-M-002-009
  • National Health Research Institutes in Taiwan. Grant Number: NHRI-EX100-10020EC

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Keywords:

  • three-dimensional imaging;
  • refractive index;
  • interferometry;
  • digital holographic microscopy;
  • image reconstruction

Abstract

Quantification of three-dimensional (3D) refractive index (RI) with sub-cellular resolution is achieved by digital holographic microtomography (DHμT) using quantitative phase images measured at multiple illumination angles. The DHμT system achieves sensitive and fast phase measurements based on iterative phase extraction algorithm and asynchronous phase shifting interferometry without any phase monitoring or active control mechanism. A reconstruction algorithm, optical diffraction tomography with projection on convex sets and total variation minimization, is implemented to substantially reduce the number of angular scattered fields needed for reconstruction without sacrificing the accuracy and quality of the reconstructed 3D RI distribution. Tomogram of a living CA9-22 cell is presented to demonstrate the performance of the method. Further, a statistical analysis of the average RI of the nucleoli, the nucleus excluding the nucleoli and the cytoplasm of twenty CA9-22 cells is performed. (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

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