CellTrace™ Calcein Blue, AM - Special Packaging

Citations & References (6)

Invitrogen™

CellTrace™ Calcein Blue, AM - Special Packaging

CellTrace calcein blue AM is a proven short-term, blue-fluorescent tracer for cell labeling. The probe possesses acetoxymethyl (AM) esters thatRead more

Catalog Number	Quantity
C34853	20 x 50 μg

Catalog number C34853

Price (MXN)

Quantity:

20 x 50 μg

CellTrace calcein blue AM is a proven short-term, blue-fluorescent tracer for cell labeling. The probe possesses acetoxymethyl (AM) esters that allow its passive diffusion across cell membranes. Upon cleavage of the AM esters by intracellular esterases, the molecule that remains (absorption/emission maxima ∼360/449 nm) is relatively polar and is retained by cells for several hours.

View a selection guide for all Nonfixable Viability Dyes for Flow Cytometry.

For Research Use Only. Not for use in diagnostic procedures.

Specifications

Detection MethodFluorescence

Dye Typecalcein blue

EmissionUV

Excitation Wavelength Range323⁄439

For Use With (Equipment)Fluorescence Microscope, Flow Cytometer

FormSolid

Product LineCellTrace

Quantity20 x 50 μg

Reagent TypeCell Tracker Compounds, Cell Labeling Reagents

Shipping ConditionRoom Temperature

SolubilityDMSO (Dimethylsulfoxide)

Label TypeOther Labels or Dyes

Product TypeViability Indicator

Unit SizeEach

Contents & Storage

Contains 20 vials of calcein blue (50 μg per vial). Store in freezer (-5 to -30°C).

Frequently asked questions (FAQs)

I need a general cytoplasmic stain that does not overlap with the GFP in my cells. What do you recommend?

Calcein AM, a green dye, is typically used as a general cytoplasmic stain, but not recommended with GFP-positive cells. For GFP-expressing cells there are other colors available: Calcein Blue AM, Calcein Violet AM, and Calcein Red-Orange AM. The retention time of these dyes in live cells is dependent upon the inherent properties of the cell.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I stained two populations of cells, one with CellTracker Green and the other with CellTracker Red, but it looks like there may be crossover of the red dye to the green cells. What is going on?

One possibility is that there is spectral bleedthrough between the dyes. Be sure to check the single-color samples by imaging the red cells in green and imaging the green cells in red, using the optimal imaging settings for the other color. If you see bleedthrough with these controls, then you will have to reduce the dye label concentration to reduce the brightness of the dyes, or choose dyes that are farther apart spectrally. If the issue isn’t bleedthrough, another possibility is that the cells were not adequately washed after staining, allowing some unincorporated dye to remain and label the other cells after they were introduced. Extending washes and wash times should help with this.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I stained my cells with Calcein, AM, but the signal went away after I fixed my cells. Why is this?

Calcein, AM diffuses into cells, the 'AM' moiety is cleaved by cellular esterases, and then the dye molecules are observed in the cytoplasm without binding to anything. This gives a 'whole cell' stain. It also means that the dyes are not crosslinked with aldehyde-based fixation and thus will be lost upon fixation. Additionally, any disruption of plasma membrane, such as with detergents or trypsinization, will lead to leakage of the dye from the cell.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I'm trying to stain my cells with CellTracker dyes or CFDA SE, but I'm not seeing much signal. What can I do?

First, make sure you aren’t staining in the presence of serum, since serum can have esterase activity that can prematurely cleave the AM group on these dyes, preventing entry into cells. After staining, it’s okay to return the cells to medium containing serum. After this, you can try increasing the concentration and label time to get a higher intensity.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I like how calcein dyes label the whole cell. How long can I track my cells with them, and can I fix them?

Calcein dyes diffuse into cells, the 'AM' moiety is cleaved by cellular esterases and then are observed in the cytoplasm without binding to anything. This provides a 'whole cell' label. Calcein dyes may be pumped out by normal cellular efflux mechanisms, sometimes within a very short time, especially for cell types that may exhibit drug resistance, unless the efflux is inhibited (such as with probenecid). The dyes are not crosslinked with aldehyde-based fixation, unlike protein-binding CellTracker dyes, and thus will be lost upon fixation. Additionally, any disruption of plasma membrane, such as with detergents or trypsinization, will lead to leakage of the dyes from the cell.

Find additional tips, troubleshooting help, and resources within our Cell Tracing and Tracking Support Center.

View all CellTrace™ Calcein Blue, AM - Special Packaging FAQs

Citations & References (6)

Citations & References

Abstract

Whole animal cell sorting of Drosophila embryos.

Authors:Krasnow MA, Cumberledge S, Manning G, Herzenberg LA, Nolan GP

Journal:Science

PubMed ID:1898782

'Use of primary culture cells has been limited by the inability to purify most types of cells, particularly cells from early developmental stages. In whole animal cell sorting (WACS), live cells derived from animals harboring a lacZ transgene are purified according to their level of beta-galactosidase expression with a fluorogenic ... More

Multiplexed labeling of viable cells for high-throughput analysis of glycine receptor function using flow cytometry.

Authors:Gilbert DF, Wilson JC, Nink V, Lynch JW, Osborne GW,

Journal:Cytometry A

PubMed ID:19184990

Flow cytometry is an important drug discovery tool because it permits high-content multiparameter analysis of individual cells. A new method dramatically enhanced screening throughput by multiplexing many discrete fixed cell populations; however, this method is not suited to assays requiring functional cellular responses. HEK293 cells were transfected with unique mutant ... More

UVA-induced apoptosis studied by the new apo/necro-Comet-assay which distinguishes viable, apoptotic and necrotic cells.

Authors:Morley N, Rapp A, Dittmar H, Salter L, Gould D, Greulich KO, Curnow A,

Journal:Mutagenesis

PubMed ID:16500949

An adaptation of the Comet-assay was developed which enables the discrimination of viable, apoptotic and necrotic single cells by use of the common Annexin-V staining and a dye exclusion test on the cells already embedded in agarose gel on glass slides. Membrane integrity (Ethidium-Homodimer exclusion), cellular esterase activity (Calcein blue-AM) ... More

Early intermediates in HIV-1 envelope glycoprotein-mediated fusion triggered by CD4 and co-receptor complexes.

Authors:Dimitrov AS, Xiao X, Dimitrov DS, Blumenthal R

Journal:J Biol Chem

PubMed ID:11397808

An early step in the process of HIV-1 entry into target cells is the activation of its envelope glycoprotein (GP120-GP41) to a fusogenic state upon binding to target cell CD4 and cognate co-receptor. Incubation of human immunodeficiency virus (HIV)-1 Env-expressing cells with an excess of CD4 and co-recepeptor-bearing target cells ... More

RANK ligand-induced elevation of cytosolic Ca2+ accelerates nuclear translocation of nuclear factor kappa B in osteoclasts.

Authors:Komarova SV, Pilkington MF, Weidema AF, Dixon SJ, Sims SM

Journal:J Biol Chem

PubMed ID:12496256

RANK ligand (RANKL) induces activation of NFkappaB, enhancing the formation, resorptive activity, and survival of osteoclasts. Ca(2+) transduces many signaling events, however, it is not known whether the actions of RANKL involve Ca(2+) signaling. We investigated the effects of RANKL on rat osteoclasts using microspectrofluorimetry and patch clamp. RANKL induced ... More

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