96-well plate for a microplate assay

Microplate assays provide information on entire cell populations rather than tracking the behavior of individual cells. This high-throughput screening analysis can provide information for whole cells, disrupted cells, or cell lysates.

Microplate assays use various cellular functions to determine viability. Metabolic activity is commonly used as a viability indicator using the reducing power of living cells (alamarBlue and PrestoBlue) or cellular redox potential (MTT and XTT). For some applications, it can be important to assess viability independent of metabolic state, such as through the release of enzymatic proteins (CyQUANT Cytotoxicity assays) or loss of membrane integrity (LIVE/DEAD kits).

Microplate assays for cell viability selection guide

The alamarBlue and PrestoBlue Cell Viability Reagents are ready-to-use resazurin-based solution that function as a cell health indicator by using the reducing power of living cells to quantitatively measure viability on microplate readers.

 alamarBlue Cell Viability ReagentalamarBlue HS Cell Viability ReagentPrestoBlue Cell Viability ReagentPrestoBlue HS Cell Viability Reagent
Assay type
  • Fluorescent
Use
  • Recommended in cases of extended viability studies or when using a high cell density in microplate assays
  • Recommended for quick viability determination in microplate assays (10-minute incubation)
Measurement
  • Detecting metabolic activity of mammalian cells, bacteria, plant, and fungi
  • Only live cells detected
  • Detecting metabolic activity of mammalian cells
  • Only live cells detected
Mechanism of detection
  • Resazurin is converted to fluorescent resorufin
Assay sensitivity
Cat. No.DAL1025A50100A13261P50200
 Learn more about differences in alamarBlue and alamarBlue HSLearn more about differences in PrestoBlue and PrestoBlue HS

CyQUANT MTT and CyQUANT XTT Cell Viability Assays are reagents that function as a cell health indicator by using the cellular redox potential of live cells to quantitively measure cell viability and proliferation on microplate readers.

 CyQUANT MTT Cell Viability AssayCyQUANT XTT Cell Viability Assay
Assay type
  • Colorimetric
Use
  • Well-established, end-point assay for quantitatively determining viability and proliferation
  • Quick and continuous assay for quantitatively determining viability and proliferation
Measurement
  • Detecting metabolic activity using the redox potential in active, live mammalian cells only
Mechanism of detection
  • Water-soluble MTT is converted into an insoluble purple formazan product
  • Water-soluble XTT is converted into a water-soluble orange formazan product
Kit components
  • MTT reagent (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)
  • Sodium Dodecyl Sulfate (SDS)
  • XTT Reagent (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide)
  • Electron Coupling Reagent
Assay sensitivity
Cat. No.V13154X12223
 Learn more about differences in CyQUANT XTT and MTT Assays for Cell Viability

CyQUANT Cytotoxicity Assays use colorimetric- or fluorescent-based detection to measure the release of cytosolic enzymes, LDH or G6PD, from damaged cells on microplate readers.

 

CyQUANT LDH Cytotoxicity Assay

CyQUANT LDH Cytotoxicity Assay- FluorescenceCyQUANT Cytotoxicity Assay Kit (G6PD Release Assay)
Assay Type
  • Colorimetric
  • Fluorescent
Use
  • Multiplex with additional cell health assays
  • Quantitate lactate dehydrogenase (LDH) release from damaged cells
  • Detection of low cell numbers
  • Quantitate number of dead cells
Measurement
  • Detection of extracellular LDH from damaged cells
  • Only damaged cells detected
  • Detection of the cytosolic enzyme glucose-6-phosphate dehydrogenase (G6PD) from damaged cells
  • Only damaged cells detected
Mechanism of detection
  • Tetrazolium salt (INT) is converted to a red formazan
  • Resazurin is converted to a red fluorescent resorufin
Cat. No.C20300C20302V23111
 Learn more about the differences in CyQUANT Cytotoxicity Assays

LIVE/DEAD kits can be used on microplate readers to discriminate live from dead cells through the influx of fluorescent dyes.

 LIVE/DEAD Viability/Cytotoxicity Kit, for mammalian cellsHCS LIVE/DEAD Green Kit
 
Assay type
  • Two-color, fluorescent assay
Use
  • Detection of live and dead cells
  • Detection of all cells and dead cells
Measurement
  • Influx of cell-impermeant dye in dead cells that have lost plasma membrane integrity
  • Influx of cell-permeant dye in live cells that reacts with intracellular esterases
  • Influx of cell-impermeant dye in dead cells that have lost plasma membrane integrity
  • Influx of cell-permeant dye(s) for total cell demarcation
Detection reagent (Ex./Em.)
  • Ethidium homodimer-1 (528/617 nm)
  • Calcium AM (488/535 nm)
  • Image-iT DEAD Green (488/515 nm)
  • NuclearMask Deep Red* (636/686 nm)
  • Hoechst 33342* (350/461 nm)

*For total cell demarcation, choose from one of these. Both are provided in the kit.
Cat. No.L3224H10290
 Learn more about LIVE/DEAD Cell Viability Assays

Microplate reader assays can be used to detect bacterial or yeast viability through the influx of fluorescent dyes.

 LIVE/DEAD BacLight Bacterial Viability KitLIVE/DEAD Yeast Viability Kit
Assay type
  • Two-color fluorescent bacterial cell viability assay
  • Two-color fluorescent yeast cell viability assay
Measurement
  • Influx of cell-impermeant dye in cells that have lost plasma membrane integrity
Readout
  • Membrane-permeant DNA dye stains intact, live cells green
  • Membrane-impermeant DNA stains dead cells red
  • Live cell determination can also be combined with fluorescent Gram staining
  • A membrane stain detects both live and dead cells with a blue signal
  • In viable cells, the vacuole stains orange/red
Detection reagent (Ex./Em.)
  • Propidium iodide (485/530 nm)
  • SYTO 9 (630 nm)
  • FUN 1 (485/530 nm)
  • Calcofluor White (620 nm)
Cat. No.L7012L7009
 Learn more about LIVE/DEAD Cell Viability Assays

The alamarBlue and PrestoBlue Cell Viability Reagents are ready-to-use resazurin-based solution that function as a cell health indicator by using the reducing power of living cells to quantitatively measure viability on microplate readers.

 alamarBlue Cell Viability ReagentalamarBlue HS Cell Viability ReagentPrestoBlue Cell Viability ReagentPrestoBlue HS Cell Viability Reagent
Assay type
  • Fluorescent
Use
  • Recommended in cases of extended viability studies or when using a high cell density in microplate assays
  • Recommended for quick viability determination in microplate assays (10-minute incubation)
Measurement
  • Detecting metabolic activity of mammalian cells, bacteria, plant, and fungi
  • Only live cells detected
  • Detecting metabolic activity of mammalian cells
  • Only live cells detected
Mechanism of detection
  • Resazurin is converted to fluorescent resorufin
Assay sensitivity
Cat. No.DAL1025A50100A13261P50200
 Learn more about differences in alamarBlue and alamarBlue HSLearn more about differences in PrestoBlue and PrestoBlue HS

CyQUANT MTT and CyQUANT XTT Cell Viability Assays are reagents that function as a cell health indicator by using the cellular redox potential of live cells to quantitively measure cell viability and proliferation on microplate readers.

 CyQUANT MTT Cell Viability AssayCyQUANT XTT Cell Viability Assay
Assay type
  • Colorimetric
Use
  • Well-established, end-point assay for quantitatively determining viability and proliferation
  • Quick and continuous assay for quantitatively determining viability and proliferation
Measurement
  • Detecting metabolic activity using the redox potential in active, live mammalian cells only
Mechanism of detection
  • Water-soluble MTT is converted into an insoluble purple formazan product
  • Water-soluble XTT is converted into a water-soluble orange formazan product
Kit components
  • MTT reagent (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)
  • Sodium Dodecyl Sulfate (SDS)
  • XTT Reagent (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide)
  • Electron Coupling Reagent
Assay sensitivity
Cat. No.V13154X12223
 Learn more about differences in CyQUANT XTT and MTT Assays for Cell Viability

CyQUANT Cytotoxicity Assays use colorimetric- or fluorescent-based detection to measure the release of cytosolic enzymes, LDH or G6PD, from damaged cells on microplate readers.

 

CyQUANT LDH Cytotoxicity Assay

CyQUANT LDH Cytotoxicity Assay- FluorescenceCyQUANT Cytotoxicity Assay Kit (G6PD Release Assay)
Assay Type
  • Colorimetric
  • Fluorescent
Use
  • Multiplex with additional cell health assays
  • Quantitate lactate dehydrogenase (LDH) release from damaged cells
  • Detection of low cell numbers
  • Quantitate number of dead cells
Measurement
  • Detection of extracellular LDH from damaged cells
  • Only damaged cells detected
  • Detection of the cytosolic enzyme glucose-6-phosphate dehydrogenase (G6PD) from damaged cells
  • Only damaged cells detected
Mechanism of detection
  • Tetrazolium salt (INT) is converted to a red formazan
  • Resazurin is converted to a red fluorescent resorufin
Cat. No.C20300C20302V23111
 Learn more about the differences in CyQUANT Cytotoxicity Assays

LIVE/DEAD kits can be used on microplate readers to discriminate live from dead cells through the influx of fluorescent dyes.

 LIVE/DEAD Viability/Cytotoxicity Kit, for mammalian cellsHCS LIVE/DEAD Green Kit
 
Assay type
  • Two-color, fluorescent assay
Use
  • Detection of live and dead cells
  • Detection of all cells and dead cells
Measurement
  • Influx of cell-impermeant dye in dead cells that have lost plasma membrane integrity
  • Influx of cell-permeant dye in live cells that reacts with intracellular esterases
  • Influx of cell-impermeant dye in dead cells that have lost plasma membrane integrity
  • Influx of cell-permeant dye(s) for total cell demarcation
Detection reagent (Ex./Em.)
  • Ethidium homodimer-1 (528/617 nm)
  • Calcium AM (488/535 nm)
  • Image-iT DEAD Green (488/515 nm)
  • NuclearMask Deep Red* (636/686 nm)
  • Hoechst 33342* (350/461 nm)

*For total cell demarcation, choose from one of these. Both are provided in the kit.
Cat. No.L3224H10290
 Learn more about LIVE/DEAD Cell Viability Assays

Microplate reader assays can be used to detect bacterial or yeast viability through the influx of fluorescent dyes.

 LIVE/DEAD BacLight Bacterial Viability KitLIVE/DEAD Yeast Viability Kit
Assay type
  • Two-color fluorescent bacterial cell viability assay
  • Two-color fluorescent yeast cell viability assay
Measurement
  • Influx of cell-impermeant dye in cells that have lost plasma membrane integrity
Readout
  • Membrane-permeant DNA dye stains intact, live cells green
  • Membrane-impermeant DNA stains dead cells red
  • Live cell determination can also be combined with fluorescent Gram staining
  • A membrane stain detects both live and dead cells with a blue signal
  • In viable cells, the vacuole stains orange/red
Detection reagent (Ex./Em.)
  • Propidium iodide (485/530 nm)
  • SYTO 9 (630 nm)
  • FUN 1 (485/530 nm)
  • Calcofluor White (620 nm)
Cat. No.L7012L7009
 Learn more about LIVE/DEAD Cell Viability Assays

Expertly detect fluorescence with Thermo Scientific plate readers

Thermo Scientific plate readers

High-sensitivity fluorescence detection for 96-1,536 samples can be quickly performed on Fluoroskan or Fluoroskan FL Microplate Fluorometer or Varioskan LUX Multimode Microplate Reader using Invitrogen reagents for optimal detection. Take advantage of automatic dynamic range selection to get optimal gain settings for each individual well and automation capabilities for even higher throughput.

Compare readers

Resources

Cell Analysis Learning Center—Find educational resources such as application notes, webinars, videos, articles, and more that cover the use of many of our reagents and kits for cell analysis.

Fluorescence SpectraViewer—Online tool for visualization of the excitation and emission of fluorescent reagents; allows for checking spectral compatibility for multiple fluorophores.

Application notes

 Application note: Evaluation of the toxic effects of PFOS in hESCs differentiating into cardiomyocytes

 Application note: Monitoring cell health with alamarBlue and PrestoBlue reagents using the Varioskan LUX Multimode Microplate Reader

BioProbes article

An Absorbance-Based Assay for Cell Health and Proliferation

Poster

 New and improved cellular health evaluation of 2D and 3D cellular models using microplate reader assays

Support

Cell Analysis Support Center—Find technical information, tips and tricks, and answers to everyday problems.