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LIVE/DEAD Cell Viability AssaysEvaluate viability in cells, bacteria, fungi, and yeast |
Thermo Fisher Scientific LIVE/DEAD cell viability assays provide for easy and sensitive differentiation of live, dead, and cytotoxic populations using fluorescence microscopy, flow cytometry, or microplate readers. These fluorescent-based LIVE/DEAD assays can be used to evaluate viability in cellular populations, bacteria, fungi, and yeast. The fluorescent dyes used in these viability assays range from blue to near-IR emission. Some viability assays permit fixation, which enables intracellular staining and neutralization of pathogens.
LIVE/DEAD cell viability kits offer 2- or 3-color discrimination of live from dead cell populations based on membrane integrity, esterase activity, metabolic activity, or structural segmentation.
Learn more about these LIVE/DEAD cell viability kits.
Platform* | Fluorescent dyes | Ex/Em (nm) ** | Fluorescent Color | Cat. No. | |
---|---|---|---|---|---|
LIVE/DEAD Viability/Cytotoxicity Kit | FC, FM, M | Calcein AM ethidium homodimer-1 | 494/517 517/617 | Green (Live) Red (Dead) | L3224 |
The LIVE/DEAD Viability/Cytotoxicity Assay Kit (Green/Deep Red) | FM, M | Calcein AM, SYTOX Deep Red | 494/517 660/682 | Green (Live) Deep Red (Dead) | L32250 |
LIVE/DEAD Cell-Mediated Cytotoxicity Kit | FC, FM, M | DiOC19(3) propidium iodide | 484/501 536/617 | Green (Live) Red (Dead) | L7010 |
LIVE/DEAD Sperm Viability Kit | FC, FM | SYBR 14 dye propidium iodide | 485/517 536/617 | Green (Live) Red (Dead) | L7011 |
LIVE/DEAD Reduced Biohazard Cell Viability Kit #1 | FC, FM, M | SYTO 10 green ethidium homodimer-2 | 484/505 535/624 | Green (Live) Red (Dead) | L7013 |
LIVE/DEAD Cell Viability Assay Kit, C12 Resazurin/SYTOX Green | FC, FM, M | SYTOX Green dye C12-resazurin | 488/530 488/575 | Green (Dead) Red (Live) | L34951 |
LIVE/DEAD Violet Viability/Vitality Kit | FM | Calcein violet AM aqua-fluorescent reactive dye | 400/452 367/528 | Violet (Live) Violet (Dead) | L34958 |
LIVE/DEAD Cell Imaging Kit | FM | Calcein AM BOBO-3 Iodide | 488/515 570/602 | Green (Live) Red (Dead) | R37601 |
HCS LIVE/DEAD Green Kit | FM, M | Hoechst 33342 Image-iT DEAD Green HCS NuclearMask Deep Red | 350/461 488/515 638/686 | Blue (DNA) Green (Dead) Red (Live) | H10290 |
*FC = flow cytometry; FM = fluorescence microscopy; M = microplate assay **Ex=excitation; Em=emission |
LIVE/DEAD cell viability kits offer 2- or 3-color discrimination of live from dead cell populations based on membrane integrity, esterase activity, metabolic activity, or structural segmentation. These viability kits been have optimized for use on fluorescence microscopy, flow cytometry, and microplate readers.
LIVE/DEAD Viability/Cytotoxicity Kit is a quick and easy 2-color viability assay based on membrane integrity and esterase activity (Figure 1). Plasma membrane integrity is determined by ethidium homodimer-1 which enters cells with compromised plasma membranes to bind DNA and emit a red fluorescence; live cells are identified by Calcein AM, a fluorogenic cell-permeant dye that is converted to a green fluorescence after interaction with intracellular esterases.
Figure 1. Live and ethanol-killed bovine pulmonary artery epithelial cells stained with the reagents in our LIVE/DEAD Cell Viability/Cytotoxicity Assay Kit. A mixture of live and ethanol-killed bovine pulmonary artery epithelial cells stained with the reagents in our LIVE/DEAD Cell Viability/Cytotoxicity Assay Kit. Live cells fluoresce bright green, whereas dead cells with compromised membranes fluoresce red-orange.
LIVE/DEAD Cell Viability Assay Kit, C12 Resazurin/SYTOX Green is a simple, 2-color viability assay to identify metabolically active cells from dead and damaged cells (Figure 2). In metabolically active cells, C12-resazurin is reduced to C12-resorufin to emit a red fluorescence. C12-resazurin is the lipophilic version of resazurin that is more permeable to live cells and the C12-resorufin product is better retained in cells. The SYTOX Green dye enters the compromised plasma membrane of dead and dying cells, and binds DNA, resulting in a green fluorescence. Cells that have undergone damage, will display a reduced red and green fluorescence.
Figure 2. LIVE/DEAD Cell Vitality Kit. Jurkat cells (T-cell leukemia, human) were induced with 10 μM camptothecin for 4 hours at 37°C, 5% CO2. The cells were incubated with the reagents in the LIVE/DEAD Cell Vitality Assay Kit and analyzed by flow cytometry. The dot plot of SYTOX Green fluorescence vs. resorufin fluorescence shows resolution of live, injured, and dead cell populations.
LIVE/DEAD kits for bacterial viability are convenient and easy-to-use 2-color fluorescent assays to discriminate between live and dead bacterial cell populations based on membrane integrity.
Learn more about these LIVE/DEAD bacterial viability kits.
Platform* | Fluorescent dyes | Ex/Em (nm) ** | Fluorescent Color | Cat. No. | |
---|---|---|---|---|---|
LIVE/DEAD BacLight Bacterial Viability Kit | FC, FM, M | SYTO 9 propidium iodide | 480/500 490/635 | Green (Live) Red (Dead) | L7007 |
LIVE/DEAD BacLight Bacterial Viability Kit, for microscopy & quantitative assays | FC, FM, M | SYTO 9 propidium iodide | 480/500 490/635 | Green (Live) Red (Dead) | L7012 |
LIVE/DEAD BacLight Bacterial Viability Kit | FC, FM, M | SYTO 9 propidium iodide | 480/500 490/635 | Green (Live) Red (Dead) | L13152 |
LIVE/DEAD BacLight Bacterial Viability and Counting Kit | FC | SYTO 9 propidium iodide | 485/498 535/617 | Green (Live) Red (Dead) | L34856 |
FilmTracer LIVE/DEAD Biofilm Viability Kit | FM | SYTO 9 propidium iodide | 482/500 490/635 | Green (Live) Red (Dead) | L10316 |
*FC = flow cytometry; FM = fluorescence microscopy; M = microplate assay **Ex=excitation; Em=emission |
LIVE/DEAD kits for bacterial viability are convenient and easy-to-use 2-color fluorescent assays to discriminate between live and dead bacterial cell populations based on membrane integrity. These kits contain SYTO 9 green-fluorescent stain and PI red-fluorescent stain that differ in their ability to enter bacteria. When used alone, SYTO 9 will stain all bacteria in a population regardless of membrane integrity; however, PI will only enter bacteria with a compromised membrane causing reduced SYTO 9 fluorescence. With an optimized mixture, dead bacteria will fluoresce red, while live bacteria will fluoresce green. These assays have been optimized for use on fluorescence microscopy (Figure 3), flow cytometry (Figure 4), and microplate readers.
Learn more: bacterial viability assays for flow cytometry
Figure 3. LIVE/DEAD BacLight Bacterial Viability Kit.Micrococcus luteus and Bacillus cereus stained with the LIVE/DEAD BacLight Bacterial Viability Kit. When incubated with the SYTO 9 stain and the propidium iodide nucleic acid stain provided in this kit, live bacteria with intact cell membranes fluoresce green and dead bacteria with compromised membranes fluoresce red.
Figure 4. Analysis of bacterial cultures using the LIVE/DEAD BacLight Bacterial Viability and Counting Kit. Suspensions of live (untreated) and dead (alcohol-treated) Staphylococcus aureus (Panels A and C) and Escherichia coli (Panels B and D) were stained and analyzed by flow cytometry according to the LIVE/DEAD BacLight Bacterial Viability and Counting Kit protocol. Using green or red fluorescence versus side scatter cytogram (Panels A and B), the bacterial populations and the bead populations were gated (left and right boxes, respectively). Events in the bacteria region of each cytogram are also displayed in red fluorescence versus green fluorescence cytograms (Panels C and D). Live and dead bacteria/mL can be calculated from either the fluorescence versus side scatter cytogram or the green fluorescence versus red fluorescence cytogram, depending on which one shows the best separation of the live and dead populations. The position of the live and dead populations in these cytograms may be dependent on cell type and gram character. Some samples may exhibit events that fall outside the defined regions and should be evaluated appropriately (e.g., see Panel D).
LIVE/DEAD kits for fungi and yeast viability offer 2-color discrimination of live and dead fungal populations based on membrane integrity or metabolic activity and fungal cell wall labeling.
Learn more about these LIVE/DEAD fungi and yeast viability kits.
Platform* | Fluorescent dyes | Ex/Em (nm) ** | Fluorescent Color | Cat. No. | |
---|---|---|---|---|---|
LIVE/DEAD Yeast Viability Kit | FM, M | FUN 1 Calcofluor White M2R | 488/530 488/560-610 365/440 | Green (All) Red-orange (Live) Blue (All) | L7009 |
LIVE/DEAD FungaLight Yeast Viability Kit | FC | SYTO9 dye propidium iodide | 485/530 485/630 | Green (Live) Red (Dead) | L34952 |
*FC = flow cytometry; FM = fluorescence microscopy; M = microplate assay **Ex=excitation; Em=emission |
LIVE/DEAD Yeast Viability Kits provide easy, reliable, and quantitative discrimination in pure or mixed cultures. Each kit offers 2-color discrimination based on membrane integrity or metabolic activity, and fungal cell wall labeling.
Learn more: yeast viability assays for flow cytometry
The Yeast Viability Kit (Figure 5) contains FUN 1 dye, that uses the plasma membrane integrity and metabolic activity of live populations to convert the yellow-green-fluorescent intracellular staining to red-orange intravacuolar structures, and Calcofluor White M2R which labels cell-wall chitin with a blue-fluorescence.
Figure 5. LIVE/DEAD Yeast Viability Kit.Saccharomyces cerevisiae stained with the FUN 1 cell stain, which generates red-fluorescent intravacuolar structures, and with Calcofluor White M2R, a blue-fluorescent cell wall stain, both components of the LIVE/DEAD Yeast Viability Kit.
The LIVE/DEAD FungaLight Yeast Viability Kit uses 2 nucleic acids stains - SYTO9 green-fluorescent stain and PI red-fluorescent stain (Figure 6). When used alone, SYTO 9 will stain all yeast in the population; while PI only enters yeast with a damaged membrane resulting in reduced SYTO 9 fluorescence. As a result, live yeast with uncompromised membrane will fluoresce green whereas dead yeast with compromised membrane will fluoresce red.
Figure 6. Saccharomyces spp. cell suspensions stained with SYTO 9 dye and propidium iodide and analyzed using a BD FACSCalibur flow cytometry system (Becton Dickinson and Co.). Panel A shows the dot plot of forward scatter vs. side scatter of an untreated Saccharomyces culture, washed and stained with SYTO 9 dye and propidium iodide as described in the LIVE/DEAD FungaLight Yeast Viability Kit protocol. The region R1 contains particles of the appropriate size for yeast cells; the forward scatter trigger is set to exclude debris in the sample. Panel B shows the R1-gated staining pattern obtained following analysis of a sample of yeast containing a mixture of both live and heat-killed cells.
LIVE/DEAD cell viability kits offer 2- or 3-color discrimination of live from dead cell populations based on membrane integrity, esterase activity, metabolic activity, or structural segmentation.
Learn more about these LIVE/DEAD cell viability kits.
Platform* | Fluorescent dyes | Ex/Em (nm) ** | Fluorescent Color | Cat. No. | |
---|---|---|---|---|---|
LIVE/DEAD Viability/Cytotoxicity Kit | FC, FM, M | Calcein AM ethidium homodimer-1 | 494/517 517/617 | Green (Live) Red (Dead) | L3224 |
The LIVE/DEAD Viability/Cytotoxicity Assay Kit (Green/Deep Red) | FM, M | Calcein AM, SYTOX Deep Red | 494/517 660/682 | Green (Live) Deep Red (Dead) | L32250 |
LIVE/DEAD Cell-Mediated Cytotoxicity Kit | FC, FM, M | DiOC19(3) propidium iodide | 484/501 536/617 | Green (Live) Red (Dead) | L7010 |
LIVE/DEAD Sperm Viability Kit | FC, FM | SYBR 14 dye propidium iodide | 485/517 536/617 | Green (Live) Red (Dead) | L7011 |
LIVE/DEAD Reduced Biohazard Cell Viability Kit #1 | FC, FM, M | SYTO 10 green ethidium homodimer-2 | 484/505 535/624 | Green (Live) Red (Dead) | L7013 |
LIVE/DEAD Cell Viability Assay Kit, C12 Resazurin/SYTOX Green | FC, FM, M | SYTOX Green dye C12-resazurin | 488/530 488/575 | Green (Dead) Red (Live) | L34951 |
LIVE/DEAD Violet Viability/Vitality Kit | FM | Calcein violet AM aqua-fluorescent reactive dye | 400/452 367/528 | Violet (Live) Violet (Dead) | L34958 |
LIVE/DEAD Cell Imaging Kit | FM | Calcein AM BOBO-3 Iodide | 488/515 570/602 | Green (Live) Red (Dead) | R37601 |
HCS LIVE/DEAD Green Kit | FM, M | Hoechst 33342 Image-iT DEAD Green HCS NuclearMask Deep Red | 350/461 488/515 638/686 | Blue (DNA) Green (Dead) Red (Live) | H10290 |
*FC = flow cytometry; FM = fluorescence microscopy; M = microplate assay **Ex=excitation; Em=emission |
LIVE/DEAD cell viability kits offer 2- or 3-color discrimination of live from dead cell populations based on membrane integrity, esterase activity, metabolic activity, or structural segmentation. These viability kits been have optimized for use on fluorescence microscopy, flow cytometry, and microplate readers.
LIVE/DEAD Viability/Cytotoxicity Kit is a quick and easy 2-color viability assay based on membrane integrity and esterase activity (Figure 1). Plasma membrane integrity is determined by ethidium homodimer-1 which enters cells with compromised plasma membranes to bind DNA and emit a red fluorescence; live cells are identified by Calcein AM, a fluorogenic cell-permeant dye that is converted to a green fluorescence after interaction with intracellular esterases.
Figure 1. Live and ethanol-killed bovine pulmonary artery epithelial cells stained with the reagents in our LIVE/DEAD Cell Viability/Cytotoxicity Assay Kit. A mixture of live and ethanol-killed bovine pulmonary artery epithelial cells stained with the reagents in our LIVE/DEAD Cell Viability/Cytotoxicity Assay Kit. Live cells fluoresce bright green, whereas dead cells with compromised membranes fluoresce red-orange.
LIVE/DEAD Cell Viability Assay Kit, C12 Resazurin/SYTOX Green is a simple, 2-color viability assay to identify metabolically active cells from dead and damaged cells (Figure 2). In metabolically active cells, C12-resazurin is reduced to C12-resorufin to emit a red fluorescence. C12-resazurin is the lipophilic version of resazurin that is more permeable to live cells and the C12-resorufin product is better retained in cells. The SYTOX Green dye enters the compromised plasma membrane of dead and dying cells, and binds DNA, resulting in a green fluorescence. Cells that have undergone damage, will display a reduced red and green fluorescence.
Figure 2. LIVE/DEAD Cell Vitality Kit. Jurkat cells (T-cell leukemia, human) were induced with 10 μM camptothecin for 4 hours at 37°C, 5% CO2. The cells were incubated with the reagents in the LIVE/DEAD Cell Vitality Assay Kit and analyzed by flow cytometry. The dot plot of SYTOX Green fluorescence vs. resorufin fluorescence shows resolution of live, injured, and dead cell populations.
LIVE/DEAD kits for bacterial viability are convenient and easy-to-use 2-color fluorescent assays to discriminate between live and dead bacterial cell populations based on membrane integrity.
Learn more about these LIVE/DEAD bacterial viability kits.
Platform* | Fluorescent dyes | Ex/Em (nm) ** | Fluorescent Color | Cat. No. | |
---|---|---|---|---|---|
LIVE/DEAD BacLight Bacterial Viability Kit | FC, FM, M | SYTO 9 propidium iodide | 480/500 490/635 | Green (Live) Red (Dead) | L7007 |
LIVE/DEAD BacLight Bacterial Viability Kit, for microscopy & quantitative assays | FC, FM, M | SYTO 9 propidium iodide | 480/500 490/635 | Green (Live) Red (Dead) | L7012 |
LIVE/DEAD BacLight Bacterial Viability Kit | FC, FM, M | SYTO 9 propidium iodide | 480/500 490/635 | Green (Live) Red (Dead) | L13152 |
LIVE/DEAD BacLight Bacterial Viability and Counting Kit | FC | SYTO 9 propidium iodide | 485/498 535/617 | Green (Live) Red (Dead) | L34856 |
FilmTracer LIVE/DEAD Biofilm Viability Kit | FM | SYTO 9 propidium iodide | 482/500 490/635 | Green (Live) Red (Dead) | L10316 |
*FC = flow cytometry; FM = fluorescence microscopy; M = microplate assay **Ex=excitation; Em=emission |
LIVE/DEAD kits for bacterial viability are convenient and easy-to-use 2-color fluorescent assays to discriminate between live and dead bacterial cell populations based on membrane integrity. These kits contain SYTO 9 green-fluorescent stain and PI red-fluorescent stain that differ in their ability to enter bacteria. When used alone, SYTO 9 will stain all bacteria in a population regardless of membrane integrity; however, PI will only enter bacteria with a compromised membrane causing reduced SYTO 9 fluorescence. With an optimized mixture, dead bacteria will fluoresce red, while live bacteria will fluoresce green. These assays have been optimized for use on fluorescence microscopy (Figure 3), flow cytometry (Figure 4), and microplate readers.
Learn more: bacterial viability assays for flow cytometry
Figure 3. LIVE/DEAD BacLight Bacterial Viability Kit.Micrococcus luteus and Bacillus cereus stained with the LIVE/DEAD BacLight Bacterial Viability Kit. When incubated with the SYTO 9 stain and the propidium iodide nucleic acid stain provided in this kit, live bacteria with intact cell membranes fluoresce green and dead bacteria with compromised membranes fluoresce red.
Figure 4. Analysis of bacterial cultures using the LIVE/DEAD BacLight Bacterial Viability and Counting Kit. Suspensions of live (untreated) and dead (alcohol-treated) Staphylococcus aureus (Panels A and C) and Escherichia coli (Panels B and D) were stained and analyzed by flow cytometry according to the LIVE/DEAD BacLight Bacterial Viability and Counting Kit protocol. Using green or red fluorescence versus side scatter cytogram (Panels A and B), the bacterial populations and the bead populations were gated (left and right boxes, respectively). Events in the bacteria region of each cytogram are also displayed in red fluorescence versus green fluorescence cytograms (Panels C and D). Live and dead bacteria/mL can be calculated from either the fluorescence versus side scatter cytogram or the green fluorescence versus red fluorescence cytogram, depending on which one shows the best separation of the live and dead populations. The position of the live and dead populations in these cytograms may be dependent on cell type and gram character. Some samples may exhibit events that fall outside the defined regions and should be evaluated appropriately (e.g., see Panel D).
LIVE/DEAD kits for fungi and yeast viability offer 2-color discrimination of live and dead fungal populations based on membrane integrity or metabolic activity and fungal cell wall labeling.
Learn more about these LIVE/DEAD fungi and yeast viability kits.
Platform* | Fluorescent dyes | Ex/Em (nm) ** | Fluorescent Color | Cat. No. | |
---|---|---|---|---|---|
LIVE/DEAD Yeast Viability Kit | FM, M | FUN 1 Calcofluor White M2R | 488/530 488/560-610 365/440 | Green (All) Red-orange (Live) Blue (All) | L7009 |
LIVE/DEAD FungaLight Yeast Viability Kit | FC | SYTO9 dye propidium iodide | 485/530 485/630 | Green (Live) Red (Dead) | L34952 |
*FC = flow cytometry; FM = fluorescence microscopy; M = microplate assay **Ex=excitation; Em=emission |
LIVE/DEAD Yeast Viability Kits provide easy, reliable, and quantitative discrimination in pure or mixed cultures. Each kit offers 2-color discrimination based on membrane integrity or metabolic activity, and fungal cell wall labeling.
Learn more: yeast viability assays for flow cytometry
The Yeast Viability Kit (Figure 5) contains FUN 1 dye, that uses the plasma membrane integrity and metabolic activity of live populations to convert the yellow-green-fluorescent intracellular staining to red-orange intravacuolar structures, and Calcofluor White M2R which labels cell-wall chitin with a blue-fluorescence.
Figure 5. LIVE/DEAD Yeast Viability Kit.Saccharomyces cerevisiae stained with the FUN 1 cell stain, which generates red-fluorescent intravacuolar structures, and with Calcofluor White M2R, a blue-fluorescent cell wall stain, both components of the LIVE/DEAD Yeast Viability Kit.
The LIVE/DEAD FungaLight Yeast Viability Kit uses 2 nucleic acids stains - SYTO9 green-fluorescent stain and PI red-fluorescent stain (Figure 6). When used alone, SYTO 9 will stain all yeast in the population; while PI only enters yeast with a damaged membrane resulting in reduced SYTO 9 fluorescence. As a result, live yeast with uncompromised membrane will fluoresce green whereas dead yeast with compromised membrane will fluoresce red.
Figure 6. Saccharomyces spp. cell suspensions stained with SYTO 9 dye and propidium iodide and analyzed using a BD FACSCalibur flow cytometry system (Becton Dickinson and Co.). Panel A shows the dot plot of forward scatter vs. side scatter of an untreated Saccharomyces culture, washed and stained with SYTO 9 dye and propidium iodide as described in the LIVE/DEAD FungaLight Yeast Viability Kit protocol. The region R1 contains particles of the appropriate size for yeast cells; the forward scatter trigger is set to exclude debris in the sample. Panel B shows the R1-gated staining pattern obtained following analysis of a sample of yeast containing a mixture of both live and heat-killed cells.
Supports all levels of experimental complexity, use the tool to compare excitation and emission spectra of fluorophores and reagents.
Protocols that fit your needs in flow cytometry ranging from sample preparation to numerous cell stimulation conditions, staining, immunophenotyping, and data analysis strategies.
Protocols that fit your needs in imaging ranging from sample and assay preparation to staining, labeling, and data analysis strategies.
Flow Cytometry Panel Builder – Design your flow cytometry panel with this online tool for a simplified, customizable experience to fit your needs
Cell Viability and Function Information – Learn about assays and techniques for evaluating cell health and function using various detection methods.
Cell Analysis Learning Center – Access educational resources to learn about applications, techniques, and basic principles during experiment planning and execution.
Cell Analysis Support Center – Find technical information, tips and tricks, and answers to everyday problems.
For Research Use Only. Not for use in diagnostic procedures.