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 nuclei
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.
Application Notes:
- Automated Kinetic Imaging Assay of Cell Proliferation in 384-Well Format
- Automated Scoring of Acid-fast Bacteria (AFB) by Fluorescent Smear Microscopy
- Automated Hemocytometer-Based Live/Dead Cell Counting using Phase Contrast and Color Brightfield Imaging
- Normalization of Agilent Seahorse XF Data by In-situ Cell Counting Using a BioTek Cytation 5
- Live Cell Imaging of Apoptosis and Necrosis
- Automated Cell Counting with High Contrast Brightfield and EVE Counting Slides
- Analysis of Nuclear Stained Cells - Using the Cytation™3 Cell Imaging Multi-Mode Microplate Reader with DAPI-Stained Cells
Infographic: How to Calculate Cell Count/Volume Using Automated Label Free Image Analysis
Related Applications:
Visual Abstracts:
BioTek’s Visual Abstracts are brief animated presentations describing the Augmented Microscopy workflow of a specific application.
Related Products:
- Lionheart FX Automated Microscope
- Lionheart LX Automated Microscope
- BioSpa Live Cell Analysis System
- Cytation 7 Cell Imaging Multi-Mode Reader
- Cytation 5 Cell Imaging Multi-Mode Reader
- Cytation 1 Cell Imaging Multi-Mode Reader
Learn more about Cell Counting applications - search our Application Notes and technical documentation in Resources.