Metrolab Blog

Cell Counting

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. BioTek’s imaging and microscopy products can count cells using a number of microscopy methods including fluorescence microscopy using fluorometric probes that intercalate with nuclear DNA, color brightfield for trypan blue-based viability measurements and label-free cell counting using either phase contrast or high contrast brightfield. Below are examples of fluorescence and stained cell counting applications using BioTek products:

Tissue Culture

  • Standard hemocytometer-based cell counting using color brightfield to count trypan blue-stained non-viable cells
  • Automated cell counting using C-chips and fluorescence microscopy
  • High throughput automated cell counting using microplates and fluorescence microscopy

Cell-based Assays

  • Cell proliferation assays using fluorescence microscopy
  • Cytotoxicity and apoptosis assays using fluorescence microscopy

Normalization of Cell-based Assays

  • Total cell counts and/or % viability after assay readout in the same microplate using fluorescence microscopy

MCF10A cells, Hoechst-labeled nucleiMCF10A cells, Hoechst-labeled nuclei

MCF10A cells, Hoechst-labeled nuclei masked for counting MCF10A cells, Hoechst-labeled nuclei masked for counting

With Gen5 3.0 Software, cell counting is both powerful and intuitive. Advanced image pre-processing tools like image smoothing and background flattening help obtain quantitative measurements from the acquired images with high accuracy and precision. Default cell counting settings provided in Gen5 are appropriate for the majority of standard cell counting applications, including 1-click cell counting that removes the complexity from image analysis.

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