In situ microscopic cytometry enables noninvasive viability assessment of animal cells by measuring entropy states |
| |
Authors: | Philipp Wiedemann Jean S Guez Hans B Wiegemann Florian Egner Juan C Quintana Diego Asanza‐Maldonado Marcos Filipaki Jeff Wilkesman Christian Schwiebert Jean P Cassar Pascal Dhulster Hajo Suhr |
| |
Institution: | 1. Mannheim University of Applied Sciences, Paul‐Wittsack‐Str. 10, Mannheim, Germany, D‐68163;2. telephone: +49 (0)621 292 6809;3. fax +49 (0)621 292 6420;4. Laboratoire ProBioGEM, Polytech‐Lille/IUT A, Université de Lille 1, Avenue Paul Langevin, Villeneuve d'Ascq, France;5. InVivo BioTech Services, Neuendorfstr. 24a, Hennigsdorf, Germany;6. Universit?t Konstanz, Computer and Information Science, Fach 697, Constance, Germany;7. Chemistry Department, FACYT, University of Carabobo, Valencia, Edo. Carabobo, Venezuela;8. Laboratoire LAGIS, Polytech‐Lille, Université de Lille 1, Avenue Paul Langevin, Villeneuve d'Ascq, France |
| |
Abstract: | 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. |
| |
Keywords: | |
|
|