Noninvasive time-dependent cytometry monitoring by digital holography

Manuel Kemmler; Markus Fratz; Dominik M. Giel; Norbert Saum; Albrecht Brandenburg; Christian Hoffmann

doi:10.1117/1.2804926

1 November 2007 Noninvasive time-dependent cytometry monitoring by digital holography

Manuel Kemmler, Markus Fratz, Dominik M. Giel, Norbert Saum, Albrecht Brandenburg, Christian Hoffmann

Author Affiliations +

Journal of Biomedical Optics, Vol. 12, Issue 6, 064002 (November 2007). https://doi.org/10.1117/1.2804926

Abstract

Using a digital holographic microscope setup, it is possible to measure dynamic volume changes in living cells. The cells were investigated time-dependently in transmission mode for different kinds of stimuli affecting their morphology. The measured phase shift was correlated to the cellular optical thickness, and then of the cell volume as well as the refractive index were calculated and interpreted. For the characterization of the digital holographic microscope setup, we have developed a transparent three-dimensional (3-D) reference chart that can be used as a lateral resolution chart and step-height resolution chart included in one substrate. For the monitoring of living cells, a biocompatible and autoclavable flow chamber was designed, which allows us to add, exchange, or dilute the fluid within the flow chamber. An integrated changeable coverslip enables inverse microscopic applications. Trypsinization, cell swelling and shrinking induced by osmolarity changes, and apoptosis served as model processes to elucidate the potential of the digital holographic microscopy (DHM).

©(2007) Society of Photo-Optical Instrumentation Engineers (SPIE)

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Manuel Kemmler, Markus Fratz, Dominik M. Giel, Norbert Saum, Albrecht Brandenburg, and Christian Hoffmann "Noninvasive time-dependent cytometry monitoring by digital holography," Journal of Biomedical Optics 12(6), 064002 (1 November 2007). https://doi.org/10.1117/1.2804926

Published: 1 November 2007

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KEYWORDS

Digital holography

Refractive index

Phase shifts

Cell death

Microscopes

Holography

Image segmentation

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