The Thermo Scientific alamarBlue Cell Viability Assay Reagent incorporates a redox-sensitive growth indicator to quantitatively measure cell viability and proliferation using fluorescent or colorimetric detection strategies.
The alamarBlue Reagent is a non-toxic, water-soluble resazurin dye that yields a fluorescent signal and a colorimetric change when incubated with metabolically active cells. The reagent is designed for cell viability and proliferation studies using mammalian cell lines, bacteria and fungi and is ideally suited for in vitro cytotoxicity studies. Additionally, cell viability can be monitored in real time when performing a variety of tissue engineering applications.
- High sensitivity and linearity – reproducibly detect as few as 50 cells
- Quantitative – provides accurate measurement over time
- Non-toxic to cells – less likely to interfere with normal metabolism
- Scalable – mix-and-read, homogeneous assay enhances speed while minimizing effort
- No cell lysis – ideal for use with time course experiments or post-measurement functional assays
- Stable – allows for continuous cell growth monitoring over a long period
- Flexible – can be used with primary cell lines, adherent or suspension cells, bacteria and fungi
- Versatile – allows fluorescent or colorimetric detection methods
Fluorescent plate reader for excitation at 530 to 560nm and emission detection at 590nm, or spectrophotometer plate reader capable of reading 570nm and 600nm absorbance.
- Monitor cell proliferation and cell viability for in vitro cytotoxicity studies (e.g., effects of environmental toxins, cytokines, drugs, siRNA/microRNA)
- Monitor cell viability in real time for 3D tissue engineering applications
Viable cells maintain a reducing environment within their cytoplasm. The alamarBlue Reagent is an oxidized form of redox indicator (resazurin dye) that is blue in color and non-fluorescent. When incubated with viable cells, the reagent changes color from blue to red and becomes fluorescent.
|Fluorescence and absorbance spectra of Thermo Scientific alamarBlue Cell Viability Reagent in oxidized and reduced states. The reagent (initially in the oxidized state) undergoes reduction in the presence of growing (viable) cells.
Cells are plated and incubated with reagents whose effects are being studied. The alamarBlue Cell Viability Assay Reagent is directly added to each well and incubated for a previously optimized time period at 37°C, 5% CO2 . The change in fluorescence or color is measured using fluorescence or UV-visible spectrophotometry.
|Protocol summary for Thermo Scientific alamarBlue Cell Viability Reagent.
|Optimization of cell number and incubation time for cell viability measurements.
|In vitro cytotoxicity study using Thermo Scientific alamarBlue Cell Viability Reagent. HEK-293 (left) or CHO-K1(right) cells were plated in a 96-well plate and exposed to various concentrations of the cytotoxic compounds, Etoposide and Actinomycin D, respectively. Cells were loaded with alamarBlue Reagent, incubated at 37°C in 5% CO2, and then the plates were measured at 545nm/590nm (Ex/Em) using the Thermo Scientific Varioscan Flash Multimode Plate Reader.
- Al-Nasiry S., Geusens N, Hassens M., Luyten C., and Pijnenborg R. (2006). The use of AlamarBlue assay for quantitative analysis of viability, migration and invasion of choriocarcinoma cells. Human Reproduction. 22(5):1304-9.
- Fields, RD., and Lancaster, MV. (1993). Dual attribute continuous monitoring of cell proliferation/cytotoxicity. American Biotechnology Laboratory. 11(4):48-50.
- Schoonen WG, Stevenson JC, Westerink WM, Horbach GJ. (2012). Cytotoxic effects of 109 reference compounds on rat H4IIE and human HepG2 hepatocytes. III: Mechanistic assays on oxygen consumption with MitoXpress and NAD(P)H production with Alamar Blue. Toxicol In Vitro. 26(3):511-25.
- Bhat S, Kumar A.(2012). Cell proliferation on three-dimensional chitosan-agarose-gelatin cryogel scaffolds for tissue engineering applications. J Biosci Bioeng. 2012 Aug 9.
- Peng L, Zhang R, Sun H, Chen L, Liu F, Yao C, Mouxuan D, Xiaodan J. (2012). PKH26 can transfer to host cells in vitro and vivo. Stem Cells Dev. 2012 Aug 22. [Epub ahead of print]
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