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Poisson-event-based analysis of cell proliferation.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Summers, Huw D 
Wills, John W 
Brown, M Rowan 
Rees, Paul 

Abstract

A protocol for the assessment of cell proliferation dynamics is presented. This is based on the measurement of cell division events and their subsequent analysis using Poisson probability statistics. Detailed analysis of proliferation dynamics in heterogeneous populations requires single cell resolution within a time series analysis and so is technically demanding to implement. Here, we show that by focusing on the events during which cells undergo division rather than directly on the cells themselves a simplified image acquisition and analysis protocol can be followed, which maintains single cell resolution and reports on the key metrics of cell proliferation. The technique is demonstrated using a microscope with 1.3 μm spatial resolution to track mitotic events within A549 and BEAS-2B cell lines, over a period of up to 48 h. Automated image processing of the bright field images using standard algorithms within the ImageJ software toolkit yielded 87% accurate recording of the manually identified, temporal, and spatial positions of the mitotic event series. Analysis of the statistics of the interevent times (i.e., times between observed mitoses in a field of view) showed that cell division conformed to a nonhomogeneous Poisson process in which the rate of occurrence of mitotic events, λ exponentially increased over time and provided values of the mean inter mitotic time of 21.1 ± 1.2 hours for the A549 cells and 25.0 ± 1.1 h for the BEAS-2B cells. Comparison of the mitotic event series for the BEAS-2B cell line to that predicted by random Poisson statistics indicated that temporal synchronisation of the cell division process was occurring within 70% of the population and that this could be increased to 85% through serum starvation of the cell culture.

Description

Keywords

Key terms: cell proliferation, computer-assisted image analysis, correlation studies, microscopy, mitosis, poisson distribution, Cell Line, Cell Proliferation, Cell Tracking, Humans, Image Interpretation, Computer-Assisted, Image Processing, Computer-Assisted, Mitosis, Single-Cell Analysis, Software

Journal Title

Cytometry A

Conference Name

Journal ISSN

1552-4922
1552-4930

Volume Title

87

Publisher

Wiley