The ability to stain live cells with a viability dye and preserve that staining pattern after fixation is critical for intracellular immunophenotyping. Exclusion of the dead cells from the data allows cleaner separation and identification of cell populations. The Invitrogen Molecular Probes LIVE/DEAD Fixable Dead Cell Stains are fixable viability dyes that help to ensure accurate assessment of cell viability in samples after fixation and/or permeabilization.
These flow cytometry–based kits provide you with tools that are:
- Flexible—eight single-channel colors available for UV, 405, 488, 532, 561, or 633 nm lasers in three packaging sizes to match your experiment
- Specific—live and dead cells are clearly differentiated, even after fixation, allowing easy exclusion of dead cells that can impact the accuracy of your results
- Simple—fit into almost any staining and immunophenotyping protocol
See also viability assays for fluorescence imaging and microplates
LIVE/DEAD Fixable Dead Cell Stains
LIVE/DEAD Fixable Dead Cell Stain Kits are based on the reaction of a fluorescent reactive dye with cellular proteins (amines). These dyes cannot penetrate live cell membranes, so only cell surface proteins are available to react with the dye, resulting in dim staining (Figure 1, LIVE). The reactive dye can permeate the damaged membranes of dead cells and stain both the interior and exterior amines, resulting more intense staining (Figure 1, DEAD).
The difference in fluorescence intensity between the live and dead cell populations is typically greater than 50-fold, thereby allowing complete, simultaneous discrimination between the two cell populations (Figure 2A). Additionally, the covalent attachment of the dyes to proteins allows preservation of discrimination of the live and dead cell populations following fixation with formaldehyde, commonly used for intracellular immunophenotyping and to inactivate pathogens (Figure 2B).
Figure 2. Retention of LIVE/DEAD Fixable Dead Cell Stains after fixation. The LIVE/ DEAD Fixable Aqua Dead Cell Stain Kit was used to differentially stain a mixture of live (left peak) and heat-treated Jurkat cells (right peak). Cells in (A) were not fixed; cells in (B) were fixed in 3.7% formaldehyde following staining. Samples were analyzed by flow cytometry using 405 nm excitation and ~525 nm emission.
LIVE/DEAD Fixable Blue stain | LIVE/DEAD Fixable Violet stain | LIVE/DEAD Fixable Aqua stain | LIVE/DEAD Fixable Yellow stain | ||
---|---|---|---|---|---|
Basis of assay | LIVE/DEAD Fixable Dead Cell Stain kits are fixable viability dyes that distinguish live cells from dead cells based on cell membrane integrity and access to available amines. Cells can then be fixed for intracellular antigen detection without loss of original cell staining pattern. Allows easy exclusion of dead cells from flow cytometry readout. | ||||
Readout | Dead cells are highly fluorescent and easy to distinguish from live cells, which are stained with minimal fluorescence | ||||
Laser (nm) |
UV
|
405
|
405
|
405
|
|
Ex/Em (nm) |
350/450
|
416/451
|
367/526
|
400/575
|
|
Bibliography |
Citations
|
||||
Multiplexable |
Yes
|
Yes
|
Yes
|
Yes
|
|
Dye compatibility | Cannot be used in combination with Invitrogen product line such as Indo-1 (blue), DAPI, or Hoechst 33342 | Cannot be used in combination with Invitrogen product line such as Pacific Blue dye, CellTrace Violet stain, FxCycle Violet stain, BV421 or eFluor 450 | Cannot be used in combination with Invitrogen product line such as Pacific Green dye, F2N12S apoptosis assay, AmCyan, or BV510 | Cannot be used in combination with Invitrogen product line such as Pacific Orange dye, Qdot 605, F2N12S apoptosis assay, or BV605 | |
Fixable |
Yes
|
Yes
|
Yes
|
Yes
|
|
Sample type |
Live cells
|
||||
Cat. No. | 80 tests | ||||
200 tests | |||||
400 tests |
LIVE/DEAD Fixable Green stain | LIVE/DEAD Fixable Red stain | |||
---|---|---|---|---|
Basis of assay | LIVE/DEAD Fixable Dead Cell Stain kits are fixable viability dyes that distinguish live cells from dead cells based on cell membrane integrity and access to available amines. Cells can then be fixed for intracellular antigen detection without loss of original cell staining pattern. Allows easy exclusion of dead cells from flow cytometry readout. | |||
Readout | Dead cells are highly fluorescent and easy to distinguish from live cells, which are stained with minimal fluorescence | |||
Laser (nm) |
488
|
488
|
561
|
|
Ex/Em (nm) |
495/520
|
595/615
|
||
Bibliography |
Citations
|
|||
Multiplexable |
Yes
|
Yes
|
||
Dye compatibility | Cannot be used in combination with Invitrogen product line such as FITC, Alexa Fluor 488, GFP, or CellTrace CSFE stain | Cannot be used in combination with Invitrogen product line such as R-PE, propidium iodide, SYTOX AADvanced stain, 7-AAD, PE-Alexa Fluor 610 and PE-Texas Red tandems | ||
Fixable |
Yes
|
Yes
|
||
Sample type |
Live cells
|
|||
Cat. No. | 80 tests | |||
200 tests | ||||
400 tests |
LIVE/DEAD Fixable Far Red stain | LIVE/DEAD Fixable Near-IR stain | ||
---|---|---|---|
Basis of assay | LIVE/DEAD Fixable Dead Cell Stain kits are fixable viability dyes that distinguish live cells from dead cells based on cell membrane integrity and access to available amines. Cells can then be fixed for intracellular antigen detection without loss of original cell staining pattern. Allows easy exclusion of dead cells from flow cytometry readout. | ||
Readout | Dead cells are highly fluorescent and easy to distinguish from live cells, which are stained with minimal fluorescence | ||
Laser (nm) |
633/635
|
633/635
|
|
Ex/Em (nm) |
650/665
|
750/775
|
|
Bibliography |
Citations
|
||
Multiplexable |
Yes
|
Yes
|
|
Dye compatibility | Cannot be used in combination with Invitrogen product line such as APC, Alexa Fluor 647 dye, FxCycle Far Red and CellTrace Far Red stains | Cannot be used in combination with Invitrogen product line such as APC-Alexa Fluor 750, APC-Cy 7 or Vybrant DyeCycle Ruby stain | |
Fixable |
Yes
|
Yes
|
|
Sample type |
Live cells
|
||
Cat. No. | 80 tests |
L34973
|
L34975
|
200 tests |
L10120
|
L10119
|
|
400 tests |
L34974
|
L34976
|
LIVE/DEAD Fixable Blue stain | LIVE/DEAD Fixable Violet stain | LIVE/DEAD Fixable Aqua stain | LIVE/DEAD Fixable Yellow stain | ||
---|---|---|---|---|---|
Basis of assay | LIVE/DEAD Fixable Dead Cell Stain kits are fixable viability dyes that distinguish live cells from dead cells based on cell membrane integrity and access to available amines. Cells can then be fixed for intracellular antigen detection without loss of original cell staining pattern. Allows easy exclusion of dead cells from flow cytometry readout. | ||||
Readout | Dead cells are highly fluorescent and easy to distinguish from live cells, which are stained with minimal fluorescence | ||||
Laser (nm) |
UV
|
405
|
405
|
405
|
|
Ex/Em (nm) |
350/450
|
416/451
|
367/526
|
400/575
|
|
Bibliography |
Citations
|
||||
Multiplexable |
Yes
|
Yes
|
Yes
|
Yes
|
|
Dye compatibility | Cannot be used in combination with Invitrogen product line such as Indo-1 (blue), DAPI, or Hoechst 33342 | Cannot be used in combination with Invitrogen product line such as Pacific Blue dye, CellTrace Violet stain, FxCycle Violet stain, BV421 or eFluor 450 | Cannot be used in combination with Invitrogen product line such as Pacific Green dye, F2N12S apoptosis assay, AmCyan, or BV510 | Cannot be used in combination with Invitrogen product line such as Pacific Orange dye, Qdot 605, F2N12S apoptosis assay, or BV605 | |
Fixable |
Yes
|
Yes
|
Yes
|
Yes
|
|
Sample type |
Live cells
|
||||
Cat. No. | 80 tests | ||||
200 tests | |||||
400 tests |
LIVE/DEAD Fixable Green stain | LIVE/DEAD Fixable Red stain | |||
---|---|---|---|---|
Basis of assay | LIVE/DEAD Fixable Dead Cell Stain kits are fixable viability dyes that distinguish live cells from dead cells based on cell membrane integrity and access to available amines. Cells can then be fixed for intracellular antigen detection without loss of original cell staining pattern. Allows easy exclusion of dead cells from flow cytometry readout. | |||
Readout | Dead cells are highly fluorescent and easy to distinguish from live cells, which are stained with minimal fluorescence | |||
Laser (nm) |
488
|
488
|
561
|
|
Ex/Em (nm) |
495/520
|
595/615
|
||
Bibliography |
Citations
|
|||
Multiplexable |
Yes
|
Yes
|
||
Dye compatibility | Cannot be used in combination with Invitrogen product line such as FITC, Alexa Fluor 488, GFP, or CellTrace CSFE stain | Cannot be used in combination with Invitrogen product line such as R-PE, propidium iodide, SYTOX AADvanced stain, 7-AAD, PE-Alexa Fluor 610 and PE-Texas Red tandems | ||
Fixable |
Yes
|
Yes
|
||
Sample type |
Live cells
|
|||
Cat. No. | 80 tests | |||
200 tests | ||||
400 tests |
LIVE/DEAD Fixable Far Red stain | LIVE/DEAD Fixable Near-IR stain | ||
---|---|---|---|
Basis of assay | LIVE/DEAD Fixable Dead Cell Stain kits are fixable viability dyes that distinguish live cells from dead cells based on cell membrane integrity and access to available amines. Cells can then be fixed for intracellular antigen detection without loss of original cell staining pattern. Allows easy exclusion of dead cells from flow cytometry readout. | ||
Readout | Dead cells are highly fluorescent and easy to distinguish from live cells, which are stained with minimal fluorescence | ||
Laser (nm) |
633/635
|
633/635
|
|
Ex/Em (nm) |
650/665
|
750/775
|
|
Bibliography |
Citations
|
||
Multiplexable |
Yes
|
Yes
|
|
Dye compatibility | Cannot be used in combination with Invitrogen product line such as APC, Alexa Fluor 647 dye, FxCycle Far Red and CellTrace Far Red stains | Cannot be used in combination with Invitrogen product line such as APC-Alexa Fluor 750, APC-Cy 7 or Vybrant DyeCycle Ruby stain | |
Fixable |
Yes
|
Yes
|
|
Sample type |
Live cells
|
||
Cat. No. | 80 tests |
L34973
|
L34975
|
200 tests |
L10120
|
L10119
|
|
400 tests |
L34974
|
L34976
|
LIVE/DEAD Fixable Dead Cell Stain Sampler Kit
Not sure about which dye will suit your needs, or need even more flexibility in your experiments? Try our LIVE/DEAD Fixable Dead Cell Stain Sampler Kit, which contains enough of each color stain to run 40 tests.
Compensation for LIVE/DEAD Fixable Dead Cell Stains
The Invitrogen ArC Amine Reactive Compensation Bead Kit provides a consistent, accurate, and simple-to-use technique for the setting of flow cytometry compensation when using any of the LIVE/DEAD Fixable Dead Cell Stain Kits or any amine-reactive dye. Advantages of the kit include:
- No need to heat-treat cells
- Optimized for LIVE/DEAD Fixable Dead Cell Stain Kits
- Fast and simple bead-based flow cytometry compensation
- Precious samples are not needed for setting compensation
- Accurate and consistent results
Beads from the ArC Bead Kit were stained with LIVE/DEAD Fixable Violet Dead Cell Stain Kit (V34955) and analyzed by flow cytometry. A mixture of the positive and negative beads in the ArC Bead Kit (A 10346) were stained with LIVE/DEAD Fixable Violet Dead Cell Stain Kit (V34955), washed, and analyzed using a flow cytometry equipped with a 405/7nm laser with 450/50 nm band pass filter. Histogram shows positive and negative peaks clearly distinguishable.
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