Imaging assays for viability use the same principles as population assays. Cells with compromised plasma membranes allow the influx of cell-impermeant DNA-binding dyes which fluoresce only when bound to DNA. Thus, DNA staining can be used as a viability indicator for individual cells within a population.

Find mammalian cell viability assays

See also cell viability assays for flow cytometry

Imaging Mammalian Cell Viability

  • High content, multi-parameter imaging assays
  • Scalable from single slides to microplates

Membrane integrity is a commonly measured parameter in cell viability assays. Cells with compromised membranes allow entry to otherwise cell-impermeant DNA-binding dyes, and this DNA staining serves as a viability indicator for individual cells within a population.

Multi-parameter assays can detect both live and dead cells to provide a total count. They can also provide measurement of esterase activity or mitochondrial function as a corollary to viability, providing more context to determine the status of both live and dead cells.

Kits with a choice of stains are optimized for high-content screening and allow for further multiplexing with fluorescent antibody-based detection.

 LIVE/DEAD® Cell Imaging Kit

Hep G2 cells stained using the LIVE/DEAD® Cell Imaging Kit.

Mammalian Cell Viability Assays

Cell membranes in damaged and dying cells have increased permeability and allow the influx of dyes. Combination assays measure multiple parameters to provide more context for determining the status of both live and dead cells.

Target Detects membrane permeability using fluorescent DNA binding dye(s)
Single-color assay Also measures esterase activity Two-color DNA binding assay Three-color assay for mitochondrial toxicity, cell toxicity, and total cell number
Readout Dead cells fluoresce green Active cells emit mostly green, and dead cells emit mostly red Dead cells emit green, total cells emit red or blue; measures damaged cells and total cell number Viability and cytotoxicity assay measuring membrane permeability and mitochondrial function, together with total cell count
Fluorescent label Image-iT® DEAD Green™ Calcein-AM Image-iT® DEAD Green™ Image-iT® DEAD Green™
BOBO™-3 iodide NuclearMask™ Deep Red or Hoechst 33342 MitoTracker® Orange CMTMRos & Hoechst 33342
Standard filter set FITC FITC FITC FITC
Texas Red® DAPI or Cy®5 DAPI or Texas Red®
Ex/Em (nm) 488/515 488/535 488/515 488/515
570/602 638/686 or 350/461 350/461 and 550/580
Bibliography Citations Citations
Multiplexing Yes Yes Yes Yes
Platform Imaging Imaging, high content Imaging, high content Imaging, high content
Format 1 vial/250 coverslips 1 kit/200 assays 1 kit/200 assays 1 kit/200 assays
Cat. No. I10291 R37601 H10290 H10295