The violet laser (405 nm) has become increrviceseasingly popular in flow cytometry due to its ability to increase the multiplexing capabilities of the flow cytometer. Life Technologies is the leader in providing reagents for the violet laser for the analysis of cellular processes including cell cycle analysis, cell viability and vitality, cell proliferation, and apoptosis. These reagents can be used on the violet laser channels to increase the multicolor capabilities of today’s flow cytometers. Using the violet laser for these parameters frees the other channels on the flow cytometer for immunophenotyping and other common flow applications.

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Cell Viability and Vitality

LIVE/DEAD® Fixable Dead Cell Stains

The fixable dead cell stains covalently interact with available surface amines, and are excluded from the interior of healthy cells. In contrast, for dead cells, the dyes label proteins throughout the cytoplasm, staining dead cells with at least 50-fold greater fluorescence than live cells. Because the labeling is covalent, stained cells can be aldehyde-fixed and permeabilized without losing viability discrimination, making it  ideal for researchers who want to fix samples before analysis. The ArC™ Amine Reactive Bead Kit is the perfect tool for setting up the compensation requirements of the fixable dead cell stains in your experiments.


  • Postfixation measurement of viability
  • Accurate intracellular staining measurements
  • Reliable viability measurements
Fixable Dead Cell Dyes

 

Use of fixable dead cell dyes. The reagents in the LIVE/DEAD® Fixable Aqua Dead Cell Stain Kit were used to differentially stain a mixture of live (left peak) and heat-treated Jurkat cells (right peak). The cells in (A) were not fixed; the cells in (B) were fixed in 3.7% formaldehyde following the staining reaction. Samples were analyzed by flow cytometry using 405 nm excitation and ~525 nm emission.

Product  
LIVE/DEAD® Fixable Aqua Dead Cell Stain Kitproduct details
LIVE/DEAD® Fixable Violet Dead Cell Stain Kitproduct details
LIVE/DEAD® Fixable Yellow Dead Cell Stain Kitproduct details
LIVE/DEAD® Fixable Yellow Dead Cell Stain Sampler Kitproduct details
ArC™ Bead Kit *for use with amine reactive dyes*product details
SYTOX® Blue Nucleic Acid Stain

SYTOX® Blue Dead Cell Stain is a high-affinity nucleic acid stain that easily penetrates cells with compromised plasma membranes but remains excluded from healthy cells. After brief incubation with SYTOX® Blue Dead Cell Stain, the nucleic acids of dead cells fluoresce bright blue when excited with 405 nm violet laser light. These properties make the SYTOX® Blue Dead Cell Stain a simple and quantitative single-step dead cell indicator for use with violet laser–equipped flow cytometers.


  • Alternative laser excitation for nucleic acid staining
  • Replacement for propidium iodide (PI) using the violet laser
  • Rapid and easy-to-use protocol for DNA staining
SYTOX® Blue Dead Cell Stain

Use of SYTOX® Blue Dead Cell Stain with Vybrant® DyeCycle™ Green Stain. Jurkat cells were stained with Vybrant® DyeCycle™ Green Stain, then stained with the impermeant DNA dye, SYTOX® Blue Dead Cell Stain, and analyzed by flow cytometry using 405 nm and 488 nm excitation, gating on SYTOX® Blue–negative cells to eliminate dead cells. Cell cycle was then evaluated using a histogram of Vybrant® DyeCycle™ Green staining.

Products  
SYTOX® Blue Dead Cell Stainproduct details

CellTrace™ Calcein Violet AM

CellTrace™ Calcein Violet AM is an optimal dye for detecting intracellular esterase activity using the violet laser. It is available either as a stand-alone reagent or in the LIVE/DEAD® Violet Viability/Vitality Kit, which combines CellTrace™ Calcein Violet fluorescence with the dead cell aqua–fluorescent reactive dye. This unique kit allows the simultaneous analysis of both live and dead cells using the violet laser.

 

  • Live cell stain for the violet laser
  • Determines physiological activity regardless of membrane status
  • Little to no non-specific background staining

 

Esterase Substrates


Combination of esterase substrates with dead cell dyes using mixtures of heat-treated and untreated cells. Chinese hamster ovary (CHO) cells were stained according to the protocol in the LIVE/DEAD® Violet Viability/Vitality Kit ( CellTrace™ Calcein Violet , AM and fixable violet dead cell stain). Cells were analyzed by flow cytometry using 405 nm excitation.

Product  
LIVE/DEAD® Violet Viability/Vitality Kit product details
CellTrace™ Calcein Violet, AM product details

 

CountBright™ Absolute Counting Beads

CountBright™ absolute counting beads are a calibrated suspension of microspheres that are brightly fluorescent across a wide range of excitation and emission wavelengths (UV to 635 nm excitation and 385nm emission). CountBright™ absolute counting beads are mixed with the cell sample and assayed via flow cytometry. By comparing the ratio of bead events to cell events, absolute numbers of cells in the sample can be calculated. Because CountBright™ beads are mixed in the test sample, absolute cell counts using this single-platform method are more accurate and less complicated than cell concentration determined using multiple-platform testing. CountBright™ absolute counting beads can be used with any sample type, including no-wash/lysed whole blood.
CountBright Counting Beads

Determination of live and dead cells and absolute cell number
Plot of calcein fluorescence collected with 530/30 nm bandpass filter vs. ethidium homodimer-1 fluorescence collected with 610/20 nm bandpass filter, showing clear separation of live and dead cells, as well as counting beads (C36950). A mixture of live and heat-killed Jurkat cells (human T-cell leukemia) was treated with the reagents in the LIVE/DEAD® Viability /Cytotoxicity Kit, counting beads were added, and the sample was analyzed by flow cytometry using 488 nm excitation.

Product  
CountBright™ Absolute Counting Beads product details

 

Ordering Information

Sku Name Size Price Qty
A10346 ArC™ Amine Reactive Compensation Bead Kit 1 kit USD 275.00
C34858 CellTrace™ Calcein Violet, AM, for 405 nm excitation - Special Packaging 20 x 25 µg USD 371.00
C36950 CountBright™ Absolute Counting Beads, for flow cytometry 5 mL USD 238.00
L3224 LIVE/DEAD® Viability/Cytotoxicity Kit, for mammalian cells 1 kit USD 386.00
L34955 LIVE/DEAD® Fixable Violet Dead Cell Stain Kit, for 405 nm excitation 1 kit USD 265.00
L34957 LIVE/DEAD® Fixable Aqua Dead Cell Stain Kit, for 405 nm excitation 1 kit USD 265.00
L34958 LIVE/DEAD® Violet Viability/Vitality Kit, for 405 nm excitation 1 kit USD 497.00
L34959 LIVE/DEAD® Fixable Yellow Dead Cell Stain Kit 1 kit USD 285.00
L34960 LIVE/DEAD® Fixable Dead Cell Stain Sampler Kit 1 kit USD 355.00
S34857 SYTOX® Blue Dead Cell Stain, for flow cytometry 1 mL USD 288.00
V35004 Vybrant® DyeCycle™ Green Stain 400 µL USD 369.00