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A High Throughput Direct Adherent Cell Analysis Method for Cell Cycle and Apoptosis using Celigo Imaging Cytometer

Leo L. Chan, Scott Cribbes, Sarah Kessel, Olivier Dery, Dmitry Kuksin, and Jean Qiu

Apoptosis and cell cycle play an important role in various aspects of preclinical pharmaceutical drug discovery and validation. The ability to quickly determine the cytotoxic effect of chemical compounds on cancer cells
allows researchers to efficiently identify potential drug candidates for further development in the pharmaceutical discovery pipeline. Recently, a plate-based imaging cytometry system, Celigo Imaging Cytometer, has been
used to for high-throughput fluorescence cell cycle and apoptosis analysis. In this study, we demonstrate the use of Celigo imaging cytometry for apoptosis and cell cycle detection by studying the dose response effect of
nocodazole on cell cycle and staurosporine on apoptosis. For cell cycle analysis, the cells are labeled with propidium iodide and BrdU. For apoptosis analysis, the cells are labeled with Annvexin V-PE and Hoechst 33342.
The experimental results were evaluated to validate the imaging cytometric capabilities of the Imaging Cytometry system. The plate-based imaging cytometer utilizes bright-field and three fluorescence channels (Blue,
Green, and Red) for multi-channel analysis. By utilizing the F theta lens technology, uniform bright-field image is captured for more accurate cell counting and analysis of the entire well. In addition, Celigo analysis
software is used to report numerous parameters allowing detailed fluorescence-based cell population characterization. The ability of Celigo to rapidly and cost-effectively perform plate-based fluorescent assays has the
potential of improving research efficiency, especially for adherent cells where plate-based cytometer does not require trypsinization for cell population analysis.

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