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In situ microscopic cytometry enables noninvasive viability assessment of animal cells by measuring entropy states
Author(s) -
Wiedemann Philipp,
Guez Jean S.,
Wiegemann Hans B.,
Egner Florian,
Quintana Juan C.,
AsanzaMaldonado Diego,
Filipaki Marcos,
Wilkesman Jeff,
Schwiebert Christian,
Cassar Jean P.,
Dhulster Pascal,
Suhr Hajo
Publication year - 2011
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.23252
Subject(s) - in situ , suspension culture , flow cytometry , cytometry , staining , biophysics , entropy (arrow of time) , viability assay , cell culture , cell , biology , chemistry , microbiology and biotechnology , physics , biochemistry , thermodynamics , genetics , organic chemistry
Current state of the art to determine the viability of animal cell suspension cultures is based on sampling and subsequent counting using specific staining assays. We demonstrate for the first time a noninvasive in situ imaging cytometry capable of determining the statistics of a morphologic transition during cell death in suspension cultures. To this end, we measure morphometric inhomogeneity—defined as information entropy—in cell in situ micrographs. We found that the cells are partitioned into two discrete entropy states broadened by phenotypical variability. During the normal course of a culture or by inducing cell death, we observe the transition of cells between these states. As shown by comparison with ex situ diagnostics, the entropy transition happens before or while the cytoplasmatic membrane is loosing its ability to exclude charged dyes. Therefore, measurement of morphometric inhomogeneity constitutes a noninvasive assessment of viability in real time. Biotechnol. Bioeng. 2011;108: 2884–2893. © 2011 Wiley Periodicals, Inc.