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Molecular Probes™ LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation
Description
- Used to determine the viability of cells prior to the fixation and permeabilization required for intracellular antibody staining or prior to elimination of biohazardous materials using formaldehyde fixation
- Kit has been optimized and validated for use with a UV laser flow cytometer
- Conveniently packaged in 40-test vials to help ensure the stability and performance of the dye over time
- Amine reactive dyes in solution will lose their effectiveness over a short period of time, therefore it is recommended to completely use the vial once rehydrated
- If this is not possible, aliquot the vials in small volumes and store at -80°C, avoiding freeze-thaw cycles
- LIVE/DEAD™ Fixable Dead Cell Stains are available in a wide variety of colors to meet multi-color panel needs
Optimal brightness
- The LIVE/DEAD™ Fixable Blue Stain was selected based on its fluorescent properties to provide a bright signal when excited with a UV laser
- The blue-fluorescent reactive dye has an excitation maximum of ∼350 nm, making it ideal for use with an UV laser, and an emission of ∼450 nm
- By switching viability staining from a channel off of the blue 488 nm laser to the UV laser, a channel is freed up on the flow cytometer for other reagents that are difficult to find in colors other than FITC or PE
- By using the UV laser, there is no spectral overlap issues with other common dyes, thus eliminating the need to compensate the viability stains from other stains in the panel
- Since live and dead cells can be discriminated using a single dye and a single channel of a flow cytometer, it is an ideal choice for multi-color experiments
Specifications
Specifications
| Cell Permeability | Impermeant |
| Cell Type | Eukaryotic Cells |
| Color | Blue |
| Content And Storage | Contains 2 vials of LIVE/DEAD™ fixable dead cell stain and 500 μL DMSO. Store at -20°C. |
| Description | LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation |
| Detection Method | Fluorescence |
| For Use With (Application) | Viability Assay |
| For Use With (Equipment) | Flow Cytometer |
| Product Type | Dead Cell Stain Kit |
| Dye Type | LIVE/DEAD™ Fixable Blue Dead Cell Stain |
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Frequently Asked Questions (FAQs)
There are two easy options. One is to heat-inactivate the cells by placing at 60 degrees C for 20 minutes. The second is to subject the cells to 70% ethanol. Alcohol-fixed cells can be stored indefinitely in the freezer until use, potentially up to several years.
Centrifuge cells, pellet, and remove supernatant.
Fix cells: Add 10 mL ice cold 70% ETOH to a 15 mL tube containing the cell pellet, adding dropwise at first while vortexing, mix well.
Store in freezer until use.
When ready to use, wash twice and resuspend in buffer of choice.
The LIVE/DEAD Fixable kits for flow cytometry analysis are compatible with fixation. These kits use amine-reactive cell-impermeant dyes that stain the cell surface of live cells and also the cytosol of dead cells-live cells are dim and dead cells are bright. Since the dye is covalently bound to the cells, it will be retained after fixation. Unfortunately, this method does not work well for imaging-based assays, as all cells are stained and it is difficult to distinguish bright dead cells from dim live cells with a microscope. Ethidium monoazide (EMA; Cat No. E1374) is a cell impermeant nucleic acid stain that can be applied to live cultures and stains only dead cells. After incubation and washing away unbound dye, the cells can be exposed to light to photoactivate EMA to crosslink to dead cell DNA. After crosslinking to dead cell DNA, the samples may be fixed and permeabilized.
Image-IT DEAD Green Viability Stain (Cat. No. I10291) for imaging and high-content screening (HCS) analysis is a live-cell impermeant DNA binding dye that is compatible with fixation and permeabilization with good retention up to 48 hours.
We also have a LIVE/DEAD Reduced Biohazard Cell Viability Kit (Cat. No. L7013) for imaging and flow analysis that contains two DNA binding dyes, SYTO 10 and Dead Red, that are sufficiently retained to be analyzed soon after 4% glutaraldehyde fixation.
Note: In general, DNA-binding dyes and calcein AM are not compatible with fixation, as these dyes are not covalently bound to components of the cell and will thus slowly diffuse out of cells after fixation, gradually staining all cells as dead.
Safety and Handling
For Research Use Only. Not for use in diagnostic procedures.
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