Hep G2 cells stained using the LIVE/DEAD Cell Imaging Kit.
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.
Imaging mammalian cell viability
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
|
Resources
Learn more from the Molecular Probes Handbook
BioProbes articles
Related Products
For Research Use Only. Not for use in diagnostic procedures.