Imaging Mammalian Cell Viability

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

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

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

  • Quick and convenient ReadyProbes Assays
  • Multi-parameter imaging assays
  • Scalable building blocks for HCS

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.

Stains with differential membrane permeability can be used to build multi-parameter assays that detect both live and dead cells to provide a total count. Multiplex assays can be used to provide additional measures of viability, such as esterase activity, metabolic activity, or mitochondrial function, providing more context than a single-parameter readout.

We offer functional reagents in a choice of color options so that you can construct and optimize multi-parameter assays for high-content analysis, including further multiplexing with fluorescent antibody-based detection.


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 assays to detect dead cells
Two-color assays to detect dead cells and total cells
Readout
Dead cells stain green
Dead cells stain red
Dead cells stain green, live cells appear blue
Dead cells stain red, total cells appear blue
Fluorescent label
DNA stain
DNA stain
DNA Stain
Propidium iodide
Hoechst 33342
Hoechst 33342
Standard filter set
FITC
Cy5
FITC
TRITC/RFP
DAPI
DAPI
Ex/Em (nm)
504/523
642/661
504/523
535/617
360/460
360/460
Multiplexing
Yes
Yes
Yes
Yes
Protocol
Microscopy
Microscopy
Microscopy
Microscopy
Format
6 x 2.5 mL dropper bottle
6 x 2.5 mL dropper bottle
Cat. No.
R37109
R37113
R37609
R37610
 
LIVE/DEAD Viability/Cytotoxicity Kit, for mammalian cells
LIVE/DEAD Cell Imaging Kit (488/570)
LIVE/DEAD Sperm Viability Kit
Target
Detects membrane permeability using fluorescent DNA- binding dye(s)
Also measures esterase activity
Measures live and membrane-compromised sperm cells
Readout
Active cells emit mostly green, and dead cells emit mostly red
Active sperm cells emit green and dead cells emit mostly red
Active sperm cells emit green and dead cells emit mostly red
Fluorescent label
Calcein AM
Calcein AM
SYBR 14 stain
Ethidium homodimer
BOBO-3 iodide
Propidium iodide
Standard filter set
FITC
FITC
FITC
TRITC/RFP
Texas Red
TRITC/RFP
Ex/Em (nm)
488/535
488/535
489/519
528/617
570/602
535/617
Multiplexing
Yes
Yes
Yes
Protocol
Microscopy
Microscopy
Microscopy
Format
1 kit/1,000 assays
1 kit/200 assays
1 kit/200–1,000 assays
Cat. No.
L3224
R37601
L7011
 
Propidium iodide
LIVE/DEAD Green Kit
Mitochondrial Health Kit
Target
Detects membrane permeability using fluorescent DNA-binding dye(s)
Readout
Single-color assay components for multiplexing
Two-color DNA binding assay
Three-color assay for mitochondrial toxicity, cell toxicity, and total cell number
Dead cells fluoresce
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
Propidium iodide
Image-iT DEAD Green
Image-iT DEAD Green
Standard filter set
FITC
TRITC/RFP
NuclearMask Deep Red or Hoechst 33342
MitoTracker Orange CMTMRos & Hoechst 33342
Ex/Em (nm)
488/515
535/617
FITC
FITC
DAPI
DAPI or Cy5
TRITC
488/515
488/515
638/686 or 350/461
350/461 and 550/580
Multiplexing
Yes
Yes
Yes
Yes
Protocol
HCS
 
Microscopy
Microscopy
Format
1 vial/250 coverslips
100 mg
1 kit/200 assays
1 kit/200 assays
Cat. No.
I10291
P1304MP
H10290
H10295
 
Target
Detects membrane permeability using fluorescent DNA-binding dye(s)
Single-color assays to detect dead cells
Two-color assays to detect dead cells and total cells
Readout
Dead cells stain green
Dead cells stain red
Dead cells stain green, live cells appear blue
Dead cells stain red, total cells appear blue
Fluorescent label
DNA stain
DNA stain
DNA Stain
Propidium iodide
Hoechst 33342
Hoechst 33342
Standard filter set
FITC
Cy5
FITC
TRITC/RFP
DAPI
DAPI
Ex/Em (nm)
504/523
642/661
504/523
535/617
360/460
360/460
Multiplexing
Yes
Yes
Yes
Yes
Protocol
Microscopy
Microscopy
Microscopy
Microscopy
Format
6 x 2.5 mL dropper bottle
6 x 2.5 mL dropper bottle
Cat. No.
R37109
R37113
R37609
R37610
 
LIVE/DEAD Viability/Cytotoxicity Kit, for mammalian cells
LIVE/DEAD Cell Imaging Kit (488/570)
LIVE/DEAD Sperm Viability Kit
Target
Detects membrane permeability using fluorescent DNA- binding dye(s)
Also measures esterase activity
Measures live and membrane-compromised sperm cells
Readout
Active cells emit mostly green, and dead cells emit mostly red
Active sperm cells emit green and dead cells emit mostly red
Active sperm cells emit green and dead cells emit mostly red
Fluorescent label
Calcein AM
Calcein AM
SYBR 14 stain
Ethidium homodimer
BOBO-3 iodide
Propidium iodide
Standard filter set
FITC
FITC
FITC
TRITC/RFP
Texas Red
TRITC/RFP
Ex/Em (nm)
488/535
488/535
489/519
528/617
570/602
535/617
Multiplexing
Yes
Yes
Yes
Protocol
Microscopy
Microscopy
Microscopy
Format
1 kit/1,000 assays
1 kit/200 assays
1 kit/200–1,000 assays
Cat. No.
L3224
R37601
L7011
 
Propidium iodide
LIVE/DEAD Green Kit
Mitochondrial Health Kit
Target
Detects membrane permeability using fluorescent DNA-binding dye(s)
Readout
Single-color assay components for multiplexing
Two-color DNA binding assay
Three-color assay for mitochondrial toxicity, cell toxicity, and total cell number
Dead cells fluoresce
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
Propidium iodide
Image-iT DEAD Green
Image-iT DEAD Green
Standard filter set
FITC
TRITC/RFP
NuclearMask Deep Red or Hoechst 33342
MitoTracker Orange CMTMRos & Hoechst 33342
Ex/Em (nm)
488/515
535/617
FITC
FITC
DAPI
DAPI or Cy5
TRITC
488/515
488/515
638/686 or 350/461
350/461 and 550/580
Multiplexing
Yes
Yes
Yes
Yes
Protocol
HCS
 
Microscopy
Microscopy
Format
1 vial/250 coverslips
100 mg
1 kit/200 assays
1 kit/200 assays
Cat. No.
I10291
P1304MP
H10290
H10295