A guide to automated apoptosis detection: How to make sense of imaging flow cytometry data

• Published in: PloS one Vol. 13; no. 5; p. e0197208
• Main Authors: Pischel, Dennis, Buchbinder, Jörn H, Sundmacher, Kai, Lavrik, Inna N, Flassig, Robert J
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 16-05-2018; Public Library of Science (PLoS)
• Subjects: Algorithms; Animals; Apoptosis; Artificial intelligence; Automation; Biology and Life Sciences; Cancer; Case studies; Cell cycle; Cell death; Cell division; Classification; Classification schemes; Computer and Information Sciences; Computer applications; Data analysis; Data mining; Discrimination; Engineering; Flow; Flow cytometry; Flow Cytometry - instrumentation; Flow Cytometry - methods; Funding; Gating; Genotype & phenotype; HeLa Cells; Humans; Imaging; Immunology; Learning algorithms; Machine learning; Measurement techniques; Methods; Microscopy; Morphology; Phenotypes; Physical Sciences; Population; Research and Analysis Methods; Software; Germany
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0197208

Imaging flow cytometry is a powerful experimental technique combining the strength of microscopy and flow cytometry to enable high-throughput characterization of cell populations on a detailed microscopic scale. This approach has an increasing importance for distinguishing between different cellular phenotypes such as proliferation, cell division and cell death. In the course of undergoing these different pathways, each cell is characterized by a high amount of properties. This makes it hard to filter the most relevant information for cell state discrimination. The traditional methods for cell state discrimination rely on dye based two-dimensional gating strategies ignoring information that is hidden in the high-dimensional property space. In order to make use of the information ignored by the traditional methods, we present a simple and efficient approach to distinguish biological states within a cell population based on machine learning techniques. We demonstrate the advantages and drawbacks of filter techniques combined with different classification schemes. These techniques are illustrated with two case studies of apoptosis detection in HeLa cells. Thereby we highlight (i) the aptitude of imaging flow cytometry regarding automated, label-free cell state discrimination and (ii) pitfalls that are frequently encountered. Additionally a MATLAB script is provided, which gives further insight regarding the computational work presented in this study.