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04511 Sigma

Live/Dead Cell Double Staining Kit

suitable for fluorescence

Synonym: Staining kit for live/dead cells

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Description

Application

The Live/Dead Cell Double Staining Kit is utilized for simultaneous fluorescence staining of viable and dead cells. This kit contains calcein-AM and propidium iodide (PI) solutions, which stain viable and dead cells, respectively. Calcein-AM, acetoxymethyl ester of calcein, is highly lipophilic and cell membrane permeable. Though calcein-AM itself is not a fluorescent molecule, the calcein generated from Calcein-AM by esterase in a viable cell emits a strong green fluorescence (λex 490 nm, λem 515 nm). Therefore, calcein-AM only stains viable cells. Alternatively, the nuclei staining dye PI cannot pass through a viable cell membrane. It reaches the nucleus by passing through disordered areas of dead cell membrane, and intercalates with the DNA double helix of the cell to emit red fluorescence (λex 535 nm, λem 617 nm). Since both calcein and PI-DNA can be excited with 490 nm light, simultaneous monitoring of viable and dead cells is possible with a fluorescence microscope. Using λex 545 nm, only dead cells can be observed.

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Milli-Q® Water Purification Solutions

Kit component only

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Solution A (Calcein AM solution) 4 × 50 μL SDS    
Solution B (propidium iodide solution) 300 μL SDS    
Safety & Documentation

Safety Information

Flash Point(F) 
185 °F
Flash Point(C) 
85 °C
Protocols & Articles

Articles

Cell Viability and Proliferation Assays

Assays to measure cellular proliferation, cell viability, and cytotoxicity are commonly used to monitor the response and health of cells in culture after treatment with various stimuli. The proper ch...
Article Based on
BioFiles v6 n5, 17–21
Keywords: Cancer, Cell proliferation, Click chemistry, Enzyme-linked immunosorbent assay, Flow cytometry, Glycolysis, Immunocytochemistry, Immunohistochemistry, Microscopy, Nucleic acid denaturation, Reductions, Solvents, Western blot

Common Cell Culture Problems: Cell Clumping

Cells in suspension may attach to one another and form clumps for a variety of reasons. The most common cause of cell clumping is the presence of free DNA and cell debris in the culture medium, which...
Keywords: Adhesion, Apoptosis, Cell culture, Cell disruption, Culture media, Environmental, Flow cytometry

Common Cell Culture Problems: Cell Death

There is perhaps nothing more frustrating to a cell culturist than removing a dish from the incubator only to find that it’s full of dead cells. Getting to the bottom of the problem can be very chall...
Keywords: Antibiotics, Apoptosis, Autophagy, Buffers, Cell culture, Cell disruption, Cryopreservation, Culture media, Environmental, Enzyme activity, Filtration, Metabolism, Peptide synthesis, Reductions

Common Cell Culture Problems: Cell Line Cross-Contamination and Misidentification

A few years after scientists working at Johns Hopkins established the first human cell line using cervical epithelial cells (HeLa) from cancer patient Henrietta Lacks in the early 1950s, it was obser...
Keywords: Cancer, Cell culture, Cryopreservation, Forensic, Genetic, Phase transitions, Polymerase chain reaction

Common Cell Culture Problems: Contamination

Overview Types of Contamination Common Causes and Prevention of Contaminants    Microbial (bacteria, fungi, yeast)    Mycoplasma    Viral Contamination    Chemical Contamination General Tips and Tech...
Keywords: Antibiotics, Antifungals, Apoptosis, Buffers, Cell attachment, Cell culture, Culture media, Detergents, Eliminations, Environmental, Filtration, Fluorescent microscopy, Gene expression, Indicators, Metabolism, Microscopy, Polymerase chain reaction, Sterilizations

Common Cell Culture Problems: Poor Attachment of Adherent Cells

Optimal cell attachment in vitro requires the interaction of healthy cells with a wide range of cell-derived attachment molecules present in media, serum, and supplements. Cultureware surfaces are fr...
Keywords: Buffers, Cell attachment, Cell culture, Culture media, Decarboxylations, Environmental, Growth factors, Lipid peroxidation, Peroxidations, Precipitation, Reductions, Respiration

Common Cell Culture Problems: Precipitates

When contamination is ruled out, turbidity in cell culture media is often explained by the precipitation of metals, proteins, and other media components. Precipitates can be harmful to cell health, a...
Keywords: Antibiotics, Cell culture, Culture media, Desiccation, Evaporation, Filtration, Microscopy, Nucleic acid denaturation, Precipitation, Purification

Protocols

Common Cell Culture Problems: Poor Cell Growth

Ensuring adequate cell growth is a critical part of collecting accurate data with cell cultures. Cells can be cultured in suspension, or as a monolayer that attaches to cultureware, such as a flask, ...
Keywords: Antibiotics, Apoptosis, Buffers, Cell attachment, Cell culture, Condensations, Cryopreservation, Culture media, Evaporation, Filtration, Microscopy, Phase transitions, Reductions

Peer-Reviewed Papers
15

References

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