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Cell Counting: Label-Free

Cell counting is used as a widespread measurement in tissue culture to assess cell density for cell-based assays, as a readout in proliferation, viability and toxicity assays, and to normalize cell-based assay results for differences in cell density.

Most cell counting methods require the presence of a fluorescent nuclear label. Intercalating nuclear stains can alter the native cell properties and limit experiment duration due to cytotoxic effects. An alternative approach requires the difficult and costly endeavor of establishing stable cell lines expressing a fluorescent nuclear marker.

Given these limitations, label-free direct cell counting has considerable advantages over methods that require fluorescent labels. The High Contrast (HC) cell counting application uses modified brightfield imaging to generate a bright point of light for each cell that is easily identified and counted by Gen5™ software without the use of labels.

In-focus image of NIH3T3 cells
High contrast brightfield
High contrast brightfield
HC Cell Counting
Gen5 object masking tool readily identifies each bright spot to generate cell count.
Overlaid images with object masks
Gen5 object masking tool readily identifies each bright spot to generate cell count.
Strong linear correlation with counts derived from nuclear label-based method

Cell counting:  Label-free



Application Notes:


Related Applications:


Visual Abstracts:

BioTek’s Visual Abstracts are brief animated presentations describing the Augmented Microscopy workflow of a specific application.

Cell Proliferation
Cell Proliferation - Label-free Cell Counting


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Learn more about Label-Free Cell Counting applications - search our Application Notes and technical documentation in Resources.


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