Transcript: Viability of Fragile Hepatocytes using Image-Based Cytometry

Primary hepatocytes are frequently used in liver disease, transplantation, drug metabolism, and toxicology studies. Because the cells are very fragile, non-fluidic, image-based analysis is the preferred method for determination of hepatocyte concentration and viability. Using the Vision Cell Analyzer, bright field and dual-fluorescence imaging are combined to accurately analyze live and dead primary hepatocytes in heterogeneous samples.

In this example, the nuclear staining dyes acridine orange and propidium iodide are used to stain the hepatocytes. 20µl of stained hepatocytes are loaded into a Cellometer Counting Chamber and inserted into the Cellometer Vision Cell Analyzer. A brightfield image is viewed to optimize focus and check cell morphology.

Select the Assay from the drop-down menu, enter the Sample ID and Dilution Factor, then click Count. Cell images are captured and analyzed based on preset parameters for live and dead nucleated cells.

Live and dead cell concentration and viability are automatically reported in less than 60 seconds.

Cellometer pattern-recognition software accurately counts individual cells within clumps.

In the fluorescent counted image, live cells are outlined in green and dead cells are outlined in red.

The diameter of each cell is measured to generate a size distribution histogram.

By entering the target cell number, researchers can use the Cellometer Sample Adjustment Calculator to determine the required volume for plating.

Data tables and cell images can be easily exported for data archiving or use in presentations and publications.

More than twenty leading toxicology groups rely on the Cellometer Vision for analysis of many species of fresh and cryopreserved hepatocytes.

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