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Proceedings Paper

Coherence-controlled holographic microscopy for live-cell quantitative phase imaging
Author(s): Tomáš Slabý; Aneta Křížová; Martin Lošt'ák; Jana Čolláková; Veronika Jůzová; Pavel Veselý; Radim Chmelík
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Paper Abstract

In this paper we present coherence-controlled holographic microscopy (CCHM) and various examples of observations of living cells including combination of CCHM with fluorescence microscopy. CCHM is a novel technique of quantitative phase imaging (QPI). It is based on grating off-axis interferometer, which is fully adapted for the use of incoherent illumination. This enables high-quality QPI free from speckles and parasitic interferences and lateral resolution of classical widefield microscopes. Label-free nature of QPI makes CCHM a useful tool for long-term observations of living cells. Moreover, coherence-gating effect induced by the use of incoherent illumination enables QPI of cells even in scattering media. Combination of CCHM with common imaging techniques brings the possibility to exploit advantages of QPI while simultaneously identifying the observed structures or processes by well-established imaging methods. We used CCHM for investigation of general parameters of cell life cycles and for research of cells reactions to different treatment. Cells were also visualized in 3D collagen gel with the use of CCHM. It was found that both the cell activity and movement of the collagen fibers can be registered. The method of CCHM in combination with fluorescence microscopy was used in order to obtain complementary information about cell morphology and identify typical morphological changes associated with different types of cell death. This combination of CCHM with common imaging technique has a potential to provide new knowledge about various processes and simultaneously their confirmation by comparison with known imaging method.

Paper Details

Date Published: 11 March 2015
PDF: 4 pages
Proc. SPIE 9336, Quantitative Phase Imaging, 93360K (11 March 2015); doi: 10.1117/12.2080128
Show Author Affiliations
Tomáš Slabý, Brno Univ. of Technology (Czech Republic)
TESCAN Brno (Czech Republic)
Aneta Křížová, Brno Univ. of Technology (Czech Republic)
TESCAN Brno (Czech Republic)
Martin Lošt'ák, Brno Univ. of Technology (Czech Republic)
TESCAN Brno (Czech Republic)
Jana Čolláková, Brno Univ. of Technology (Czech Republic)
Veronika Jůzová, TESCAN Brno (Czech Republic)
Pavel Veselý, Brno Univ. of Technology (Czech Republic)
Radim Chmelík, Brno Univ. of Technology (Czech Republic)
TESCAN Brno (Czech Republic)

Published in SPIE Proceedings Vol. 9336:
Quantitative Phase Imaging
Gabriel Popescu; YongKeun Park, Editor(s)

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