Mammalian Cell Viability

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

Imaging mammalian cell viability

Quick and convenient ReadyProbes Assays

Multi-parameter imaging assays

Scalable building blocks for HCS

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

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.

	NucGreen Dead 488 ReadyProbes Reagent	NucRed Dead 647 ReadyProbes Reagent	ReadyProbes Cell Viability Imaging Kit (Blue/Green)	ReadyProbes Cell Viability Imaging Kit (Blue/Red)
Target	Detects membrane permeability using fluorescent DNA-binding dye(s)
Target	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
Fluorescent label	DNA stain	DNA stain	Hoechst 33342	Hoechst 33342
Standard filter set	FITC	Cy5	FITC	TRITC/RFP
Standard filter set	FITC	Cy5	DAPI	DAPI
Ex/Em (nm)	504/523	642/661	504/523	535/617
Ex/Em (nm)	504/523	642/661	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)
Target	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
Fluorescent label	Ethidium homodimer	BOBO-3 iodide	Propidium iodide
Standard filter set	FITC	FITC	FITC
Standard filter set	TRITC/RFP	Texas Red	TRITC/RFP
Ex/Em (nm)	488/535	488/535	489/519
Ex/Em (nm)	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

	Image-iT DEAD Green Viability Stain	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
Readout	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

ReadyProbes assays

	NucGreen Dead 488 ReadyProbes Reagent	NucRed Dead 647 ReadyProbes Reagent	ReadyProbes Cell Viability Imaging Kit (Blue/Green)	ReadyProbes Cell Viability Imaging Kit (Blue/Red)
Target	Detects membrane permeability using fluorescent DNA-binding dye(s)
Target	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
Fluorescent label	DNA stain	DNA stain	Hoechst 33342	Hoechst 33342
Standard filter set	FITC	Cy5	FITC	TRITC/RFP
Standard filter set	FITC	Cy5	DAPI	DAPI
Ex/Em (nm)	504/523	642/661	504/523	535/617
Ex/Em (nm)	504/523	642/661	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

High-performance assays

	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)
Target	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
Fluorescent label	Ethidium homodimer	BOBO-3 iodide	Propidium iodide
Standard filter set	FITC	FITC	FITC
Standard filter set	TRITC/RFP	Texas Red	TRITC/RFP
Ex/Em (nm)	488/535	488/535	489/519
Ex/Em (nm)	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

HCS assays

	Image-iT DEAD Green Viability Stain	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
Readout	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

Cytotoxicity—Assays for Cell Viability, Proliferation and Function—Chapter 15
Oxidative Stress—Probes for Reactive Oxygen Species, Including Nitric Oxide—Chapter 18
Mitochondria—Probes for Mitochondria—Section 12.2

Imaging mammalian cell viability

Mammalian cell viability assays

Resources

Learn more from the Molecular Probes Handbook

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