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Cell Meter™ BX590 fixable viability dye

Detection of Jurkat cell viability by Cell Meter™ fixable viability dye. Jurkat cells were treated and stained with Cell Meter™ BX590 (Cat#22514), and then fixed in 3.7% formaldehyde and analyzed by flow cytometry.  The dead cell population (Blue peak)  is easily distinguished from the live cell population (Red peak)  with PE-TexasRed channel, and nearly identical results were obtained before and after fixation.
Detection of Jurkat cell viability by Cell Meter™ fixable viability dye. Jurkat cells were treated and stained with Cell Meter™ BX590 (Cat#22514), and then fixed in 3.7% formaldehyde and analyzed by flow cytometry.  The dead cell population (Blue peak)  is easily distinguished from the live cell population (Red peak)  with PE-TexasRed channel, and nearly identical results were obtained before and after fixation.
Ordering information
Price ()
Catalog Number22514
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Physical properties
SolventDMSO
Spectral properties
Excitation (nm)492
Emission (nm)579
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501

OverviewpdfSDSpdfProtocol


Excitation (nm)
492
Emission (nm)
579
Discrimination and exclusion of dead cells from live cells allows cleaner separation and identification of cell populations. Cell Meter™ fixable viability dyes are a large family of cell-impermeable fluorescent viability dyes that are optimized to match the major excitation lasers of common flow cytometers, such as 350, 405, 488, 633 and 647 nm. These dyes are impermeant to live cells but permeant to cells with compromised membranes. They irreversibly react with amine- and thiol-containing proteins and other cellular components. Since dead or fixed cells with a compromised membrane more readily react with Cell Meter™ fixable cell stains, thus stain brighter than live cells with an intact membrane, these dyes can be used to assess live vs. dead status of mammalian cells. There are a few factors to be considered when using these dyes, e.g., the titration of each dye to ensure that live cells have minimal to no staining. Cell Meter™ BX590 fixable cell stain is optimized to be excited with the blue laser at 488 nm with emission at 590 nm. Compared to other commercially similar viability dyes, this fixable viability dye is much more robust and stable.

Platform


Flow cytometer

Excitation488 nm laser
Emission575/26 nm filter
Instrument specification(s)PE channel

Fluorescence microscope

ExcitationCy3/TRITC filter set
EmissionCy3/TRITC filter set
Recommended plateBlack wall/clear bottom

Example protocol


AT A GLANCE

Protocol summary
  1. Prepare samples in HHBS (0.5 mL/assay)
  2. Add Cell Meter™ BX590 to the cell suspension
  3. Stain the cells at room temperature for 20 - 60 minutes
  4. Wash the cells
  5. Fix the cells (optional)
  6. Examine the sample with flow cytometer using 575/26 nm filter (PE channel) and/or fluorescence microscope using Cy3/TRITC filter set 

Important
Thaw at room temperature before starting the experiment.

PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

Cell Meter™ BX590 stock solution (500X)
Add 200 µL of DMSO into the Cell Meter™ BX590 vial to make 500X stock solution.
Note     The unused Cell Meter™ BX590 stock solution should be divided into single use aliquots and stored at -20 °C. Avoid repeated freeze/thaw cycles.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Treat cells as desired.
  2. Wash cells once with HHBS or the azide- and serum/protein-free buffer of your choice.
  3. Resuspend cells at 5 - 10 × 10 6/mL in HHBS or in the azide- and serum/protein-free buffer of your choice.
  4. Add 1 µL of 500X Cell Meter™ BX590 stock solution to 0.5 mL of cells/assay and mix it well.
  5. Incubate at room temperature, 5% CO2 incubator for 20 - 60 minutes, protected from light.
    Note     The optimal stain concentrations and incubation time should be experimentally determined for different cell lines.
  6. Wash cells twice and resuspend cells with HHBS or the buffer of your choice.
  7. Fix cells as desired (optional).
  8. Analyze cells with a flow cytometer using 575/26 nm filter (PE channel) and/or fluorescence microscope using Cy3/TRITC filter set. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)492
Emission (nm)579

References


View all 50 references: Citation Explorer
CFSE dilution to study human T and NK cell proliferation in vitro.
Authors: Terrén, Iñigo and Orrantia, Ane and Vitallé, Joana and Zenarruzabeitia, Olatz and Borrego, Francisco
Journal: Methods in enzymology (2020): 239-255
Using Carboxy Fluorescein Succinimidyl Ester (CFSE) to Identify Quiescent Glioblastoma Stem-Like Cells.
Authors: Azari, Hassan and Deleyrolle, Loic P and Reynolds, Brent A
Journal: Methods in molecular biology (Clifton, N.J.) (2018): 59-67
Estimates and impact of lymphocyte division parameters from CFSE data using mathematical modelling.
Authors: Mazzocco, Pauline and Bernard, Samuel and Pujo-Menjouet, Laurent
Journal: PloS one (2017): e0179768
Assessment of lymphocyte proliferation for diagnostic purpose: Comparison of CFSE staining, Ki-67 expression and 3H-thymidine incorporation.
Authors: Lašťovička, Jan and Rataj, Michal and Bartůňková, Jiřina
Journal: Human immunology (2016): 1215-1222
Analysis of CFSE time-series data using division-, age- and label-structured population models.
Authors: Hross, Sabrina and Hasenauer, Jan
Journal: Bioinformatics (Oxford, England) (2016): 2321-9
Poor association of allergen-specific antibody, T- and B-cell responses revealed with recombinant allergens and a CFSE dilution-based assay.
Authors: Eckl-Dorna, J and Campana, R and Valenta, R and Niederberger, V
Journal: Allergy (2015): 1222-9
Quality control of extracorporeal photochemotherapy: Proliferation assay using CFSE validated according to ISO 15189:2007 standards.
Authors: Faivre, Lionel and Lecouflet, Lucie and Liu, Wang-Qing and Khadher, Isabelle and Lahaie, Camille and Vidal, Michel and Legouvello, Sabine and Beaumont, Jean-Louis and Bierling, Philippe and Rouard, Hélène and Birebent, Brigitte
Journal: Cytometry. Part B, Clinical cytometry (2015): 30-9
Quality control of extracorporeal photochemotherapy: Proliferation assay using CFSE validated according to ISO 15189:2007 standards.
Authors: Lionel, Faivre and Lucie, Lecouflet and Wang-Qing, Liu and Isabelle, Khadher and Camille, Lahaie and Michel, Vidal and Sabine, Legouvello and Jean-Louis, Beaumont and Philippe, Bierling and Hélène, Rouard and Brigitte, Birebent
Journal: Cytometry. Part B, Clinical cytometry (2014)
Mathematical models for CFSE labelled lymphocyte dynamics: asymmetry and time-lag in division.
Authors: Luzyanina, Tatyana and Cupovic, Jovana and Ludewig, Burkhard and Bocharov, Gennady
Journal: Journal of mathematical biology (2014): 1547-83
FlowMax: A Computational Tool for Maximum Likelihood Deconvolution of CFSE Time Courses.
Authors: Shokhirev, Maxim Nikolaievich and Hoffmann, Alexander
Journal: PloS one (2013): e67620