Multimodal label-free growth and morphology characterization of different cell types in a single culture with quantitative digital holographic phase microscopy

Björn Kemper; Jana Wibbeling; Lena Kastl; Jürgen Schnekenburger; Steffi Ketelhut

doi:10.1117/12.2079994

11 March 2015 Multimodal label-free growth and morphology characterization of different cell types in a single culture with quantitative digital holographic phase microscopy

Björn Kemper, Jana Wibbeling, Lena Kastl, Jürgen Schnekenburger, Steffi Ketelhut

Author Affiliations +

Proceedings Volume 9336, Quantitative Phase Imaging; 933617 (2015) https://doi.org/10.1117/12.2079994
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

For the analysis of the impact of pharmaceuticals or pathogens on different cellular phenotypes under identical measurement conditions and to analyze interactions between different cellular specimens a minimally-invasive quantitative observation of different cell types in a single culture is of particular interest. Digital holographic microscopy (DHM), a var-iant of quantitative phase microscopy (QPM), provides high resolution detection of optical path length changes that is suitable for stain-free minimally-invasive live cell analysis. Due to low light intensities for object illumination, QPM minimizes the interaction with the sample and has been demonstrated in particular to be suitable for long-term time-lapse investigations, e.g., for the detection of cell morphology alterations due to drugs and toxins. Furthermore, QPM has been demonstrated to be a versatile tool for the quantification of cellular growth and motility. Thus, we studied the feasibility of QPM for the analysis of mixed cell cultures and explored if quantitative phase images provide sufficient information to distinguish between different cell types and to extract cell specific parameters. For the experiments quantitative phase imaging with DHM was utilized. Mixed cell cultures with different cell types were observed with quantitative DHM phase contrast up to 35 h. The obtained series of quantitative phase images were evaluated by adapted algorithms for image segmentation. From the segmented images the area covered by the cells, the cellular dry mass and the mean cell thickness were calculated and used in the further analysis as parameters to quantify the reliability of the measurement principle. The obtained results demonstrate that it is possible to characterize the growth of cell types with different mor-phology features separately in a single culture.

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Björn Kemper, Jana Wibbeling, Lena Kastl, Jürgen Schnekenburger, and Steffi Ketelhut "Multimodal label-free growth and morphology characterization of different cell types in a single culture with quantitative digital holographic phase microscopy", Proc. SPIE 9336, Quantitative Phase Imaging, 933617 (11 March 2015); https://doi.org/10.1117/12.2079994

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