Analysis of mixed cell cultures with quantitative digital holographic phase microscopy

Björn Kemper; Jana Wibbeling; Steffi Ketelhut

doi:10.1117/12.2051310

8 May 2014 Analysis of mixed cell cultures with quantitative digital holographic phase microscopy

Björn Kemper, Jana Wibbeling, Steffi Ketelhut

Proceedings Volume 9129, Biophotonics: Photonic Solutions for Better Health Care IV; 91290W (2014) https://doi.org/10.1117/12.2051310
Event: SPIE Photonics Europe, 2014, Brussels, Belgium

Abstract

In order to study, for example, the influence of pharmaceuticals or pathogens on different cell types under identical measurement conditions and to analyze interactions between different cellular specimens a minimally-invasive quantitative observation of mixed cell cultures is of particular interest. 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 is in particular 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, the extraction morphological parameters and cell motility. We studied the feasibility of QPM for the analysis of mixed cell cultures. It was 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 digital holographic microscopy (DHM) was utilized. Mixed cell cultures with different types of human pancreatic tumor cells 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 cellular dry mass and the mean cell thickness were calculated and used in the further analysis as parameters to quantify the reliability the measurement principle. The obtained results demonstrate that it is possible to characterize the growth of cell types with different morphologies in a mixed cell culture separately by consideration of specimen size and cell thickness in the evaluation of quantitative DHM phase images.

Citation Download Citation

Björn Kemper, Jana Wibbeling, and Steffi Ketelhut "Analysis of mixed cell cultures with quantitative digital holographic phase microscopy", Proc. SPIE 9129, Biophotonics: Photonic Solutions for Better Health Care IV, 91290W (8 May 2014); https://doi.org/10.1117/12.2051310

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