LIVE/DEAD® Fixable Dead Cell Stains
|Invitrogen recognizes the importance of eliminating dead cells from your experimental analysis without compromising the integrity of your data. The Molecular Probes® LIVE/DEAD® Fixable Dead Cell Stain Kits are designed 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:
- Fixable—Staining retained after fixation for simple live/dead analysis with intracellular phenotyping
- Simple—Fits into almost any staining and phenotyping protocols
- Flexible—Seven colors from which to choose for UV, 405, 488, or 633 lasers
Choose the Right Kit
Dead Cell Stain Kit
|Blue Dead Cell Stain||UV||450||L23105|
|Violet Dead Cell Stain||405||440||L34955|
|Aqua Dead Cell Stain||405||525||L34957|
|Yellow Dead Cell Stain||405||570||L34959|
|Green Dead Cell Stain||488 or 532||530||L23101|
|Red Dead Cell Stain||488 or 561||615||L23102|
|Far Red Dead Cell Stain||633/635||665||L10120|
|Near-IR Dead Cell Stain||633/635||775||L10119|
|Dead Cell Stain Sampler||Various||450–775||L34960|
How It Works
Molecular Probes® LIVE/DEAD® Fixable Dead Cell Stain Kits are based on the reaction of a fluorescent reactive dye with cellular proteins (amines).
- Viable Cells — Because dyes can not penetrate the cell membrane, only cell surface proteins are available to reactive with dye resulting in dim staining (Figure 1).
- Dead Cells — The reactive dye can permeate damaged membranes and stain both the interior and exterior amines resulting more intense staining (Figure 1).
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 2 cell populations.
Additionally, the covalent attachment of the dyes to proteins allows preservation of the live- and dead-cell population discrimination following fixation with formaldehye, under conditions that are commonly used for intracellular immunophenotyping and to inactivate pathogens.
| Figure 1. Principle of LIVE/DEAD® Fixable Dead Cell Stain Kits.
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Live cells (top) react with the kit’s fluorescent dye only on their surface to yield weak total fluorescence. Dead cells with damaged membranes (bottom) react with the dye on their surface as well as n the cell interior, yielding bright total fluorescence.
The LIVE/DEAD® Fixable Dead Cell Stain Kits offer a broad range of advantages:
- Reliable, Post-Fixation Measurement of Viability — Common nucleic acids stains like propidium iodide (PI ) tend to leak out of cells after washing and after fixation all cells will stain positive. With the LIVE/DEAD® Fixable Dead Cell Stain Kits, the “snapshot” of viability taken prior to antibody or other reagent staining is maintained throughout the staining process, and beyond.
- Accurate Intracellular Staining Measurements — Induced cytokine and signaling responses are notoriously difficult to detect due to low stain signals, and this is complicated by nonspecific binding of reagents to dead cells.
- Removal of False Positive Dead Cells — Other methods often give false positive results as they tend to bind nonspecifically to many reagents. However, live- vs. dead-cell staining is unequivocal using LIVE/DEAD® Fixable Dead Cell Stain Kits.
- Flexibility in Experimental Design — Multiple colors for four major lasers means that these single-color assays use only one channel of one laser on a flow cytometer, leaving other channels available for more common reagents and multicolor experiments.
||Live-cell gating using the LIVE/DEAD® Fixable Violet Dead Cell Stain Kit eliminates staining artifacts from analysis. In a comparison between live-cell gating using scatter (A), and live-cell gating using LIVE/DEAD® Fixable Violet dye (B), staining artifacts using scatter gating are illustrated. The significant number of dead cells in subsequent analysis using scatter (C) are noted, as compared to the use of the LIVE/DEAD® Fixable Violet dye (D) to eliminate dead cells. Reproduced with permission from Elsevier (Amine reactive dyes: an effective tool to discriminate live and dead cells in polychromatic flow cytometry. Perfetto SP, et.al., J Immunol Methods 313:199(2006)).|
Experts agree that the ArC™ Amine Reactive Compensation Bead Kit is the perfect tools for setting up the compensation requirements of the LIVE/DEAD® Fixable Dead Cell Stains in your experiments.
|ArC™ Amine Reactive Compensation Bead Kit||1 Kit||A10346|