What you need to know

Kits for Mammalian Cells

Kit Name Platform* Fluorescent Dyes Ex/Em (nm) Em Colors SKU
LIVE/DEAD® Viability/Cytotoxicity Kit *for mammalian cells*FC, FM, Mcalcein AM ethidium homodimer-1494/517
517/617
Green (Live)
Red (Dead)
L3224
LIVE/DEAD® Cell-Mediated Cytotoxicity Kit *for animal cells* *2000 assays*FC, FM, MDiOC18(3)
propidium iodide
484/501
536/617
Green (Live)
Red (Dead)
L7010
LIVE/DEAD® Sperm Viability Kit *200-1000 assays*FC, FMSYBR® 14 dye
propidium iodide
485/517
536/617
Green (Live)
Red (Dead)
L7011
LIVE/DEAD® Cell Vitality Assay Kit *C12-resazurin/SYTOX® Green* *1000 assays*FC, FM, MSYTOX® Green dye
C12-resazurin
488/530
488/575
Green (Dead)
Red (Live)
L34951
*FC=flow cytometry; FM=fluorescence microscopy; M=microplate assay
Principle of Assay
  • LIVE/DEAD® Viability/Cytotoxicity Kit—Membrane-permeant calcein AM is cleaved by esterases in live cells to yield cytoplasmic green fluorescence, and membrane-impermeant ethidium homodimer-1 labels nucleic acids of membrane-compromised cells with red fluorescence. 

  • LIVE/DEAD® Cell-Mediated Cytotoxicity Kit—Target cells are preincubated with the green-fluorescent membrane stain DiOC18(3) and then mixed with effector cells in the presence of the red-fluorescent, membrane-impermeant dye, propidium iodide. Live and dead target cells retain their green-fluorescent membrane stain; target and effector cells with compromised membranes exhibit red-fluorescent nucleic acid staining; live effector cells are nonfluorescent.

  •  LIVE/DEAD® Sperm Viability Kit—Membrane-permeant SYBR® 14 nucleic acid stain labels live sperm with green fluorescence, and membrane-impermeant propidium iodide labels the nucleic acids of membrane-compromised sperm with red fluorescence.

  •  LIVE/DEAD® Cell Vitality Assay Kit—The assay is based on the reduction of C12-resazurin to red-fluorescent C12-resorufin in metabolically active cells and on the uptake of the cell-impermeant, green-fluorescent nucleic acid stain, SYTOX® Green dye, in cells with compromised plasma membranes (usually late apoptotic and necrotic cells). In this assay, dead cells emit mostly green fluorescence and healthy, metabolically active cells emit mostly red fluorescence; injured cells exhibit reduced red and green fluorescence.

LIVE/DEAD® Cell Viability/Cytotoxicity Assay Kit (L3224)
. A mixture of live and ethanol-killed bovine pulmonary artery epithelial cells were 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® Sperm Viability Kit (L7011)
. Live sperm with intact membranes are labeled with our proprietary cell-permeant nucleic acid stain, SYBR® 14, and fluoresce green. Dead sperm, which have been killed by unprotected freeze-thawing, are labeled with propidium iodide (P1304MP, P3566, P21493) and fluoresce red-orange. The image was contributed by Duane L. Garner, School of Veterinary Medicine, University of Nevada, Reno, and Lawrence A. Johnson, USDA Agricultural Research Service.

  LIVE/DEAD Cell Vitality Kit (L34951). 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 versusresorufin fluorescence shows resolution of live, injured, and dead cell populations.

Kits for Bacteria

Kit Name Package
Size
Platform* Fluorescent Dyes Ex/Em (nm) Em Colors SKU
LIVE/DEAD® BacLight™ Bacterial Viability KitStains are supplied in a mixed, two-component formulationFC, FM, MSYTO® 9 dye
propidium iodide
485/530
485/630
Green (Live)
Red (Dead)
L7007
LIVE/DEAD® BacLight™ Bacterial Viability KitThe stains are provided as separate solutionsFC, FM, MSYTO® 9 dye
propidium iodide
485/530
485/630
Green (Live)
Red (Dead)
L7012
LIVE/DEAD® BacLight™ Bacterial Viability KitSeparate dyes are dry and premeasured into pairs of polyethylene transfer pipetsFC, FM, MSYTO® 9 dye
propidium iodide
485/530
485/630
Green (Live)
Red (Dead)
L13152
LIVE/DEAD® BacLight Bacterial Viability and Counting KitMicrosphere standard is also includedFCSYTO® 9 dye
propidium iodide
485/530
485/630
Green (Live)
Red (Dead)
L34856
*FC=flow cytometry; FM=fluorescence microscopy; M=microplate assay
Principle of Assay
  • LIVE/DEAD® BacLight™ Bacterial Viability Kit—Membrane-permeant SYTO® 9 labels live bacteria with green fluorescence; membrane-impermeant propidium iodide labels membrane-compromised bacteria with red fluorescence.

  • LIVE/DEAD® BacLight Bacterial Viability and Counting Kit—Membrane-permeant SYTO® 9 labels live bacteria with green fluorescence; membrane-impermeant propidium iodide labels membrane-compromised bacteria with red fluorescence. The calibrated suspension of polystyrene microspheres serves as a standard for the volume of suspension analyzed and is clearly distinguishable from stained bacteria in a cytogram with fluorescence versus side scatter.

Use of our LIVE/DEAD® BacLight™ Bacterial Viability Kit
(L7007, L7012, L13152) to identify individual live and dead bacteria along a chain of Streptococcus pyogenes. This image was photographed in a single exposure through an Omega Optical triple bandpass filter set.
 
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 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).

 

Kits for Yeast and Fungi

Kit Name Platform* Fluorescent Dyes Ex/Em Em Colors SKU
LIVE/DEAD® Yeast Viability KitFM, MFUN 1
Calcofluor White M2R

488/530 (all)
488/560-610 (Live)

365/440

Green (All)
Red-orange (Live)
Blue (All)

L7009
LIVE/DEAD® FungaLight™ Yeast Viability KitFCSYTO® 9 dye
propidium iodide
485/530
485/630
Green (Live)
Red (Dead)
L34952
*FC=flow cytometry; FM=fluorescence microscopy; M=microplate assay
Principle of Assay
  • LIVE/DEAD® Yeast Viability Kit—Plasma membrane integrity and metabolic function of fungi are required to convert the yellow-green–fluorescent intracellular staining of FUN 1 into red-orange–fluorescent intravacuolar structures; Calcofluor White M2R labels cell-wall chitin with blue fluorescence regardless of metabolic state.

  • LIVE/DEAD® FungaLight™ Yeast Viability Kit—SYTO® 9 nucleic acid stain generally labels all yeast in a population – those with intact membranes and those with damaged membranes. In contrast, propidium iodide penetrates only yeast with damaged membranes, causing a reduction in the SYTO® 9 stain fluorescence by fluorescence resonance energy transfer (FRET) when both dyes are present. As a result, yeast with intact membranes stain fluorescent green, whereas yeast with damaged membranes stain fluorescent red.

  LIVE/DEAD® Yeast Viability Kit  (L7009). 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 probes are provided in our LIVE/DEAD® Yeast Viability Kit (L7009). FUN® 1 cell stain is also available separately (F7030).


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 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® Fixable Stains for Flow Cytometry

Kit Name Platform* Em (nm) Laser Line
Compatibility (nm)
SKU
LIVE/DEAD® Fixable Blue Dead Cell Stain KitFlow Cytometry450UVL23105
LIVE/DEAD® Fixable Violet Dead Cell Stain KitFlow Cytometry451405L34955
LIVE/DEAD® Fixable Aqua Dead Cell Stain KitFlow Cytometry525405L34957
LIVE/DEAD® Fixable Yellow Dead Cell Stain Kit Flow Cytometry575
405
L34959
LIVE/DEAD® Fixable Green Dead Cell Stain KitFlow Cytometry520488L23101
LIVE/DEAD® Fixable Red Dead Cell Stain KitFlow Cytometry615488L23102
LIVE/DEAD® Fixable Far Red Dead Cell Stain KitFlow Cytometry665633 or 635 L10120
LIVE/DEAD® Fixable Near-IR Dead Cell Stain KitFlow Cytometry775633 or 635 L10119
LIVE/DEAD® Fixable Dead Cell Stain Sampler KitFlow CytometryVarious
Various
L34960
Principle of Assay
  • LIVE/DEAD® Fixable Dead Cell Stain Kits—Live cells react with the fluorescent reactive dye only on their surface to yield weakly fluorescent cells. Cells with compromised membranes react with the dye throughout their volume, yielding brightly stained cells. Subsequent fixation inactivates pathogens without distorting the staining pattern.

Learn More about LIVE/DEAD® Fixable Dead Cell Stain Kits

Ordering Information