4.6 Article

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

Journal

JOURNAL OF BIOPHOTONICS
Volume 6, Issue 5, Pages 416-424

Publisher

WILEY-V C H VERLAG GMBH
DOI: 10.1002/jbio.201200022

Keywords

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

Funding

  1. National Science Council in Taiwan [NSC 98-2112-M-002-009]
  2. National Health Research Institutes in Taiwan [NHRI-EX100-10020EC]

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Quantification of three-dimensional (3D) refractive index (RI) with sub-cellular resolution is achieved by digital holographic microtomography (DHT) using quantitative phase images measured at multiple illumination angles. The DHT 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. ((c) 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

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