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Live-or-Dye™ Fixable Viability Stains for Flow Cytometry

Live-or-Dye™ Fixable Viability Staining Kits are designed for discrimination between live and dead cells during flow cytometry. Live/dead stains are useful probes to include when analyzing cell surface protein expression by flow cytometry, because they allow intracellular fluorescence signal from dead cells with permeable plasma membranes to be excluded from analysis. In microscopy, live/dead stains allow unambiguous visual discrimination of dead cells.

Live-or-Dye™ Fixable Viability Stains are cell membrane impermeable amine-reactive dyes. The dyes enter dead cells that have compromised membrane integrity and covalently label free amines on intracellular proteins. On live cells, the dyes react with surface proteins, but these are much less abundant than intracellular proteins, resulting in low staining levels compared to dead cells (Figure 1). Live-or-Dye™ Fixable Viability Staining Kits can also be used to discriminate live from dead cells during microscopy. So far four of the dyes have been validated for use in microscopy: Live-or-Dye™ 488/515, Live-or-Dye™ 568/583, Live-or-Dye™ 594/614, and Live-or-Dye™ 640/662.

Live-or-Dye™ labeling is extremely stable, allowing the cells to be fixed and permeabilized without loss of fluorescence or dye transfer between cells. The staining protocol has been optimized to maximize live/dead discrimination with minimal live cell staining, in order to prevent interference with immunostaining. Biotium offers a selection of eight amine-reactive different Live-or-Dye™ viability stains and one nuclear-labeling Live-or-Dye™ viability stain, spanning the fluorescence spectrum, for maximal flexibility in multi-color analysis.

Live-or-Dye fixable viability stains
Principle of Live-or-Dye discrimination of live and dead cells.
Live-Or-Dye Fixable Viability Stains are available in eight colors.

Live-or-Dye NucFix™ Red is a unique, cell membrane impermeable dye that specifically stains the nuclei of dead cells. Unlike other commonly used nuclear stains such as Propidium Iodide or DRAQ7, NucFix labeling is fixable.

For more information, download the Live-or-Dye Flyer and Product Information Sheets:

Live-or-Dye Flyer

Live-or-Dye Product Information Sheet

Live-or-Dye NucFix Red Product Information Sheet

live or dye nucfix red
Discrimination of live and dead cells using Live-or-Dye NucFix Red Fixable Viability Stain.
A. Live or heat-killed Jurkat cells were stained with Live-or-Dye NucFix Red. Heat killed cells (solid peak) showed much higher fluorescence intensity compared to live cells (white peak), allowing the two populations to be clearly distinguished.
B. Ethanol-treated HeLa cells were stained with NucFix Red together with Hoechst to label all cell nuclei. Killed cells show bright, red nuclear Live-or-Dye fluorescence staining, compared to no staining seen in live cells (blue Hoechst-stained nuclei).
Catalog No.Product DescriptionLaser lineEmission filterAbs/Em
32002, 32002-TLive-or-Dye™ 350/448355 nmDAPI or Violet347/448 nm
32003, 32003-TLive-or-Dye™ 405/452405 nmPacific Blue408/452 nm
32009, 32009-TLive-or-Dye™ 405/545405 nmAmCyan395/545 nm
32004, 32004-TLive-or-Dye™ 488/515488 nmFITC490/515 nm
32005, 32005-TLive-or-Dye™ 568/583488 or 561 nmPE562/583 nm
32006, 32006-TLive-or-Dye™ 594/614488 or 561 nmPE-Texas Red561/624 nm
32007, 32007-TLive-or-Dye™ 640/662633 or 640 nmAPC642/662 nm
32008, 32008-TLive-or-Dye™ 755/777633 or 640 nmAPC-Cy7755/777 nm
32010, 32010-TLive-or-Dye NucFix™ Red488 or 532 nmPE-Texas Red520/610 nm

Also see our other Products for Products for Flow Cytometry.