Cell Cycle Analysis

Cell cycle analysis is often used for the screening of compounds that affect cell growth and proliferation. The Celigo allows users to conduct cell cycle analysis on adherent cells using assays typically developed for flow cytometry. The flexible gating interface of the Celigo software is used to identify and quantify populations of cells in each phase of the cell cycle. This analysis can be conducted using a single dye such as DAPI or using a second marker for DNA synthesis such as a BrdU stain. Moreover it has the potential to be multiplexed with a marker for mitosis such as histone H3 phosphorylation. The Expression Analysis application on the Celigo provides the following benefits:

• Histogram representation of DNA content similar to flow cytometry
• Scatter plot representation of DNA content vs. DNA synthesis stain to identify each phase of the cell cycle
• Easy-to-use interface that streamlines the application from image acquisition to the generation of multi-well plate level data
• Flexibility of naming channels, segmentation features ,and cell classes to customize researchers applications

Cell cycle analysis on Celigo. A549 cells were analyzed for cell cycle phase determination using the Expression Analysis application on Celigo. Histogram representation of DNA content similar to flow cytometry (upper left). The typical “Horse Shoe” visualization of the cell cycle is achieved by plotting the integrated intensity of DNA synthesis stain vs. DNA content stain (upper right). The lower left shows a 2-color image of the BrdU and DAPI stains. The lower right shows a heat map of the plate level data results for a specific output such as percentage of cells in S-phase.

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