eFluor® Cell Proliferation Dyes
These membrane-permeable fluorescent dyes bind to cellular proteins containing primary amines; as cells divide, the dye is distributed equally between daughter cells. Cell division is therefore easily measured by flow cytometry as successive halving of the fluorescence intensity of the dye.
Cells labeled with eFluor Cell Proliferation Dyes may be fixed and permeabilized using standard formaldehyde-containing fixatives and saponin-based permeabilization buffers, allowing further analysis of intracellular targets. Each dye is named after its emission spectra to enable ease of selection for use in multicolor flow cytometry staining panels. Cell Proliferation Dyes are also used for long term tracking of labeled cells which have been introduced in vivo by adoptive transfer.
- Visualize up to 7 generations
- Use in vitro and in vivo
- Ideal for use in combination with GFP or YFP
Cell Proliferation Dye eFluor 450 - Ideal for the Violet Laser
Cell Proliferation Dye eFluor 450 - in vitro
Mouse spenocytes were labeled with 10 uM Cell Proliferation Dye eFluor 450 and cultured for 3 days with (blue histogram) or without (purple histogram) Con A. Cells were stained with Anti-Mouse CD4 PE, Anti-Mouse CD25 PerCP-Cy5.5, and 7-AAD. Viable CD4+ cells were used for analysis.
Cell Proliferation Dye eFluor 450 - in vitro
Splenocytes from Thy1.1 mice were labeled with 10 uM Cell Proliferation Dye eFluor 450 and then injected into C57BI/6 mice (purple histogram) or B6D2F1 mice (blue histogram). Splenocytes from the C57BI/6 mice or B6D2F1 mice were collected 48 hours (left) or 72 hours (right) after injection of the labeled cells. Cells were stained with Anti-Mouse CD4 APC, Anti-Mouse/Rat CD90.1 (Thy1.1) PE, Anti-Mouse CD8a PerCP-eFluor 710, and Fixable Viability Dye eFluor 780. Viable Thy1.1+CD4+ cells were used for analysis. Unlabeled C57Bi/6 or B6D2F1 cells (Thy1.1-CD4+) are shown in grey.
Cell Proliferation Dye eFluor 670 - Excited by the Red Laser
Cell Proliferation Dye eFluor 670
Mouse splenocytes were labeled with 1 uM Cell Proliferation Dye eFluor 670 and adoptively transferred into C57BI/6 mice. Three days later, cells were analyzed for cell division, measured as discrete peaks of decreasing fluorescence of dye in stimulated (pink histogram), or undivided cells with a single, bright peak (purple histogram).
Selected publications utilizing Cell Proliferation Dye eFluor 670:
- Zhou et al. J Immunol 2011; 187, 4170-4177
- Xu et al. J Immunol 2011; 186, 6597-6606
CFSE Proliferation Dye
CFSE [5-(and 6)-Carboxyfluorescein diacetate succinimidyl ester] is an established reagent for analyzing cell proliferation. The non-fluorescent CFSE dye readily crosses intact cell membranes, where intracellular esterases cleave its acetate groups to yield the fluorescent carboxyfluorescein molecule. Cell division can be measured as successive halving of the fluorescence intensity of dye with a peak excitation of 494 nm and peak emission of 521 nm.
CFSE
T-cell receptor transgenic, OVA-reactive OT-I cells undergo several rounds of cell division in response to OVA, but not PBS, in vivo. Lymph node cells from OT-I mice were labeled with 1 uM CFSE and adoptively transferred into C57BI/6 mice. Mice were then immunized with with OVA or PBS. Three days later, splenocytes were stained with Anti-Mouse/Rat Thy1.1 PerCP-Cy5.5 and Anti-Mouse CD8 eFluor 450 to gate on the OT-I cells (left). Cells in the OT-I gate (CD8+Thy1.1+) were analyzed for cell division (right), measured as discrete peaks of decreasing fluorescence of CFSE in OVA-immunized mice (purple histogram), or undivided cells with a single, bright peak in PBS-immunized mice (blue histogram).
CyTRAK Orange™ and DRAQ5™ DNA Labeling Dyes
These are membrane-permeable anthraquinone dyes with high affinity for double stranded DNA. They are membrane-permeable, and can label live or fixed/dead cells. DRAQ5™ can be used in fluorescence microscopy applications as a nuclear counterstain for distinguishing nucleated and non-nucleated cells. CyTRAK Orange™ is ideal for fluorescent microscopy; it can be used to identify and discriminate the nucleus and cytoplasm without the need for a second dye due to its high intensity staining of the nucleus and low intensity staining of the cytoplasm.
CyTRAK Orange
U2-OS human osteosarcoma cells, counterstained with CyTRAK Orange (courtesy of Biostatus).
