Invitrogen™ LIVE/DEAD™ Cell Imaging Kit (488/570)

Catalog No.	Quantity
R37601	1 mL kit

Catalog No. R37601 Supplier Invitrogen™ Supplier No. R37601

Description

Includes

Component A: 10 x 1mL vials; Component B: 1 x 10 vials, dried down

Sensitive two-color fluorescence cell viability assay optimized for FITC and Texas Red™ filters

• Allows discrimination between live and dead cells with two probes that measure recognized parameters of cytotoxicity and cell viability: intracellular esterase activity and plasma membrane integrity
• Based on cell-permeable dye for staining of live cells and cell-impermeable dye for staining of dead and dying cells
• Adapted for imaging platforms by optimizing for common green and red imaging filters used with FITC and Texas Red
• Live cell component produces intense, uniform green fluorescence in live cells (ex/em 488 nm/515nm
• Dead cell component produces predominantly nuclear red fluorescence (ex/em 570nm/602nm) in cells with compromised cell membranes, strong indicator of cell death and cytotoxicity
• Sensitive detection of key measures of cell health: viability, live/dead ratio, and cytotoxicity
• Ease of use with minimal dilutions
• Live cells are distinguished by presence of ubiquitous intracellular esterase activity as determined by the enzymatic conversion of virtually nonfluorescent cell-permeant calcein AM to intensely fluorescent calcein, which is well-retained within live cells
• Red component is cell-impermeant and therefore only enters cells with damaged membranes
• In dying and dead cells bright red fluorescence is generated upon binding to DNA
• Background fluorescence levels are inherently low with this assay technique because dyes are virtually nonfluorescent before interacting with cells

Cell Analysis, Cell Counting and Viability, Cell Counting, Viability, and Cryopreservation, Cell Culture, Cell Viability and Cytotoxicity, Cell Viability, Proliferation and Function

Order Info

Shipping Condition: Dry Ice

Specifications

• Cell Type: Mammalian Cells
• Color: Red, Green
• Content And Storage: • Component A: 10 x 1 mL vials
  • Component B: 1 x 10 vials (dried down)
  
  Store at -20°C
  
• Description: LIVE/DEAD™ Cell Imaging Kit (488/570)
• Detection Method: Fluorescence
• For Use With (Application): Viability Assay
• For Use With (Equipment): Floid Cell Imaging System, Fluorescence Microscope
• Product Type: Cell Imaging Kit
• Dye Type: Other Label(s) or Dye(s)
• Emission: 515/602
• Excitation Wavelength Range: 488, 570 nm
• Form: Frozen
• Format: Tube(s)
• Incubation Time: 15 min
• Product Line: LIVE/DEAD, Molecular Probes
• Quantity: 1 mL kit
• Shipping Condition: Dry Ice

Frequently Asked Questions (FAQs)

I need to use a dead cell control for my viability assay. Do you have a protocol for killing cells for this?

Heat killing is commonly used. Place your cells in a tube in buffer and heat at 60^oC for 20 minutes. You can also kill your cells by fixing them with ice cold 70% ethanol for 15 minutes. The ethanol-killed cells can then be stored at -20^oC until needed, at which point you wash out the ethanol and replace with buffer.

Can I use the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) with fixed cells?

The LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) is used for identifying live and dead cells using fluorescence-based staining. It cannot be used with fixed cells.

What is the composition of the Live Green (Component A) of the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601)?

The formulation of the solution as well as the amount and concentration of Calcein, AM provided in the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) is proprietary information.

Is the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) only a one-time use or is there a way to aliquot and freeze it down?

Once the Live Green (Component A) and Dead Red (Component B) are mixed, this solution should be used immediately for one-time use and should not be stored.

What is the nuclear stain in the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601)?

The nuclear stain in the LIVE/DEAD Cell Imaging Kit (488/570) is BOBO-3. The amount, concentration, and formulation is proprietary.

What cell density do you recommend using for the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601)?

As a general guideline, cell density should be within the range of 1 x 10^6 cells/mL of staining solution. Higher cell densities may result in poor staining (low signal) and lower cell densities may result in over-staining.

Could Ca++ or Mg++ ions interfere with staining using LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601)?

No, Ca++ or Mg++ ions do not interfere with staining using LIVE/DEAD Cell Imaging Kit (488/570) unless when used at high concentrations; salts at high concentrations can promote precipitation of various dyes.

How do I prepare dead cell controls for LIVE/DEAD cell viability assays?

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.

I wish to assay the viability of my cells over several days. Can I use any of your LIVE/DEAD imaging kits to do this?

No, it is not possible to assay the viability of the same population of cells longer than a few hours; you will need to use replicate samples. DNA binding dyes are toxic to cells; stained cells should be imaged as soon as possible after staining. Calcein AM is not retained in cells and may be actively effluxed out in the range from minutes to several hours, dependent upon the cell type. Calcein AM is not toxic to cells, so it can be added repeatedly to the same samples. You can assay the proliferation of the same sample of cells over several days using alamarBlue reagent or PrestoBlue reagent, as these dyes are non-toxic to cells.

Safety and Handling

WARNING: Cancer - www.P65Warnings.ca.gov