Transcript: Image-based Cell Cycle Analysis

Cellometer Vision CBA is a simple, image-based instrument optimized for analysis of cell-based assays.

For this cell cycle experiment, Jurkat cells are fixed with ethanol and incubated with a propidium iodide staining solution for 40 minutes at 37 degrees C. After centrifuging cells and resuspending in PBS, 20 µl of sample is pipetted into the Cellometer counting chamber. Insert the chamber into the Vision CBA instrument.

Select the Cell Cycle Assay from the drop-down menu. Enter the specific Sample ID, then click count.

In less than 3 minutes, the Vision CBA takes multiple cell images and analyzes the images based on pre-set parameters for the Cell Cycle Assay selected.

When cell imaging and counting is complete, the initial results table displays total cells counted, cell concentration, and mean cell diameter.

The brightfield image can be viewed to verify cell morphology. The fluorescent image shows that all cells have been stained with propidium iodide. The fluorescent counted image differentiates individual cells from cell clumps.

Click export. After defining a location for the export file, an optimized data report will be displayed. Cell cycle data is displayed as a colorized histogram showing phase population (apoptotic phase, G0/G1 phase, S phase, and G2/M phase) based on fluorescence intensity. The automatic gating can be manually adjusted. Associated data tables update automatically to reflect revised percentages.

Users can choose to print a second report containing both brightfield and fluorescent images. Data tables and cell images can be easily exported for use in presentations and publications.

After removing the disposable Cellometer cell-counting chamber, the Vision CBA is ready to analyze the next sample. No washing or instrument set-up is required.

Cellometer Vision CBA offers researchers a simple, image-based alternative to flow cytometry. Visit www.nexcelom.com or contact Nexcelom today to schedule an on-site demonstration with an experienced applications specialist.