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ViaFluor® SE Cell Proliferation Kits

ViaFluor® SE Cell Proliferation Kits use amine-reactive dyes to covalently label the cell cytoplasm, and are used to monitor cell division by flow cytometry. The dyes also can be used to stably label cells to image cell morphology, or to track cell populations in mixed co-culture experiments. The labeling withstands fixation and permeabilization for immunostaining.

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Dye Color

ViaFluor® CFSE, ViaFluor® 405, ViaFluor® 488

Cellular localization

Nucleus & cytoplasm

Apoptosis/viability marker

All cell stain

For live or fixed cells

For live/intact cells, Covalent & fixable stains

Assay type/options

Cell division tracking, Extended staining (several days to weeks), Real-time imaging

Detection method/readout

Fluorescence microscopy, Flow cytometry

Cell permeability

Membrane permeant

Fixation options

Fix after staining (formaldehyde), Fix after staining (methanol), Permeabilize after staining

Colors

Blue, Green

Dye Color
Size
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price
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Trial size (1 vial)
Set of 10 vials
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Product Description

ViaFluor®  SE Cell Proliferation Kits use amine-reactive dyes to covalently label cells throughout the cell cytoplasm and intracellular compartments. Cell proliferation dyes are commonly used to monitor cell division by flow cytometry. The dyes also can be used to stably label cells to image cell morphology, or to track cell populations in mixed co-culture experiments. Labeling is covalent, so it withstands fixation and permeabilization for subsequent immunostaining.

Features

  • Non-toxic dyes covalently label cell cytoplasm
  • Track cell proliferation in vivo or in vitro by dye dilution in flow cytometry
  • Long term imaging of cell morphology or co-cultures by microscopy
  • Staining can be fixed and permeabilized for immunofluorescence
  • Violet ViaFluor® 405 offers excellent performance at lower cost
  • Green ViaFluor® 488 SE is much less toxic than CFSE

The membrane permeable compound is non-fluorescent until it enters viable cells, where it is hydrolyzed by cytoplasmic esterase enzymes to releases the fluorescent amine-reactive dye. The dyes then covalently react with amine groups on intracellular proteins, forming fluorescent conjugates that are retained in the cell, while excess unreacted dye is washed away.  Immediately after staining, a single, bright fluorescent population will be detected by flow cytometry. With each cell division, daughter cells inherit roughly half of the fluorescent label, allowing the number of cell divisions that occur after labeling to be detected by the appearance of successively dimmer fluorescent peaks on a flow cytometry histogram compared to cells analyzed immediately after staining.  Thus, cell proliferation dyes can be used to track multiple cell divisions of cells grown in culture or injected in vivo after labeling with the ViaFluor® SE dye.

ViaFluor® SE dyes also can be used for imaging cell morphology or identifying cells in co-culture by microscopy. Because the staining is non-toxic and well-retained, it can be used for imaging  live cells over time. See our Tech Tip: Using ViaFluor® SE Stains for Cell Tracing and Co-Culture.

Kit Components:

  • 10 lyophilized dye vials (full size kits)
  • 1 lyophilized dye vial (trial size kits)
  • 1 vial of anhydrous DMSO for preparing stock solutions

When used at 1 uM in 1 mL of cells, at a cell density of 1 x 106 cells mL, each dye vial can be used for 90-100 labelings; the final number of assays that can be performed per kit depends on the dye concentration used (see product protocol under downloads).

ViaFluor® kits come with 3 dye options:

  • ViaFluor® CFSE (also known as CFDA-SE) can be detected in the FITC channel.
  • ViaFluor® 488 SE, an improved alternative to CFSE; detected in the FITC channel.
  • ViaFluor® 405 SE for the violet laser; can be detected in the Pacific Blue® channel.

ViaFluor® CFSE can be used as a cytoplasmic stain in yeast. However ViaFluor® 405 SE does not stain well in yeast.
Carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE or CFSE) is also available separately.

Please note that ViaFluor® 568 (Cat. # 30080) has been discontinued as of 12-15-17.

ProductCatalog No.Ex/Em (nm)Flow detection channelFeatures
ViaFluor® 405 Cell Proliferation Kit30068/30068-T408/452 Pacific Blue®
  • Track cell division by dye dilution using flow cytometry

  • ViaFluor® 488 is a unique, improved green dye to replace CFSE

  • ViaFluor® 405 works as well as CellTrace™ Violet
ViaFluor® 488 Cell Proliferation Kit30086/30086-T493/532FITC
ViaFluor® CFSE Cell Proliferation Kit30050495/519FITC
Pacific Blue is a registered trademark of Thermo Fisher Scientific.

 

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Support & Faq

Apoptosis, Necrosis, and Cell Viability Kits

Fixation is not recommended after staining with the Viability/Cytotoxicity Assay Kit (cat. # 30027) as the dead cell dye, EthD-III will transfer to all cells and not maintain dead cell-specific staining.

Viability qPCR (vPCR) can be an alternative strategy. Using dead cell-specific dyes, PMA (cat. # 40019) or PMAxx (cat. # 40069) that covalently modify DNA (of only dead cells) after photo-crosslinking, a simple quantitative PCR (qPCR) amplification is used to selectively amplify live-cell DNA. Learn more about the vPCR technology.

Category: Apoptosis, Necrosis, and Cell Viability Kits

← FAQs

The ViaFluor® SE dyes are susceptible to hydrolysis. Ideally the DMSO stock solution should be prepared on the day of use, with the intent that each vial supplied is for single use. If smaller aliquots are desired, it would be best to aliquot the initial DMSO stock, at the time of reconstitution, into single use quantities and store at -20C, protected from light and moisture (desiccated), for up to a month. However, activity may be reduced over time. The dyes should only be added to aqueous buffer immediately before staining. The aqueous solution cannot be stored for reuse.

Category: Apoptosis, Necrosis, and Cell Viability Kits

← FAQs

MTT and XTT are colorimetric based assays, while resazurin can be measured using colorimetric or fluorescence detection. MTT is not a soluble product, so the cells must be lysed to solubilize the formazan salt before absorbance can be measured. XTT and resazurin do not require cell lysis, allowing kinetic monitoring of the same samples at different timepoints.

Category: Apoptosis, Necrosis, and Cell Viability Kits

← FAQs

← FAQs

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