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Invitrogen™ LIVE/DEAD™ Yeast Viability Kit

Catalog No. L7009
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Catalog No. L7009 Supplier Invitrogen™ Supplier No. L7009
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Description

Viability required to change green intracelluar stain into red-orange intravacuolar structures

The LIVE/DEAD Yeast Viability Kit combines a novel two-color fluorescent probe for yeast viability, FUN™ 1, with a fluorescent fungal surface labeling reagent, Calcofluor™ White M2R. Plasma membrane integrity and metabolic function of fungi are required to convert the yellow-green-fluorescent intracellular staining of FUN 1 into red-orange intravacuolar structures; Calcofluor White M2R labels cell-wall chitin with blue-fluorescence regardless of metabolic state.

Analysis of Yeast, Cell Analysis, Cell Viability and Cytotoxicity, Cell Viability, Proliferation and Function, Microbiological Analysis, Yeast Viability and Cytotoxicity

Order Info

Shipping Condition: Room temperature

Specifications

Cell Type Yeast Cells
Color Yellow-Green, Red-Orange, Blue
Content And Storage Store in freezer at -5°C to -30°C and protect from light.
Description LIVE/DEAD™ Yeast Viability Kit
Detection Method Fluorescence
For Use With (Application) Viability Assay
For Use With (Equipment) Fluorescence Microscope, Fluorometer, Flow Cytometer, Microplate Reader
Product Type Yeast Viability Kit
Dye Type Other Label(s) or Dye(s)
Emission 530/620, 500/550
Excitation Wavelength Range 485, 450 nm
Format Tube(s), 96-well plate, Slide(s)
Product Line LIVE/DEAD
Quantity 1 kit
Shipping Condition Room Temperature
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Frequently Asked Questions (FAQs)

Which species of yeast/fungi can I use with the LIVE/DEAD Yeast Viability Kit?

The LIVE/DEAD Yeast Viability Kit has been tested on the following fungi: S. cerevisiae (five different strains), Candida pseudotropicalis, Neurospora crassa and Aspergillus nidulans. A good correlation between the results obtained with the LIVE/DEAD Yeast Viability Kit and those obtained with standard plate counts was achieved with both S. cerevisiae and C. pseudotropicalis. Tests have been performed on both logarithmically growing cultures and cells that have undergone different forms of environmental stress
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.
Can FUN 1 stained cells be examined by flow cytometry?

Yes. Use a 488 nm laser line and standard FITC and PE channels for two-color detection of green (dead/metabolically inactive cells) and red (live, metabolically active cells) emission.

Live, metabolically active fungi transport FUN 1 into vacuoles to give a red-shifted fluorescence versus green/yellow fluorescence in the nucleus and cytoplasm of dead or metabolically-inactive cells. Is this a reliable indicator of fungal viability?

No. FUN 1 accumulates into vacuoles by an unknown transport pathway, but any mutants/ recombinant cells or experimental treatments that result in a deficiency or block in vesicle-mediate transport into vacuoles may result in cells that do not have red vacuoles, even though the cells are live and metabolically active. For more information see J Microbiol Methods 78:208 (2009).

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