Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction
Abstract
Tetrazolium salts have become some of the most widely used tools in cell biology for measuring the metabolic activity of cells ranging from mammalian to microbial origin. With mammalian cells, fractionation studies indicate that the reduced pyridine nucleotide cofactor, NADH, is responsible for most MTT reduction and this is supported by studies with whole cells. MTT reduction is associated not only with mitochondria, but also with the cytoplasm and with non-mitochondrial membranes including the endosome/lysosome compartment and the plasma membrane. The net positive charge on tetrazolium salts like MTT and NBT appears to be the predominant factor involved in their cellular uptake via the plasma membrane potential. However, second generation tetrazolium dyes that form water-soluble formazans and require an intermediate electron acceptor for reduction (XTT, WST-1 and to some extent, MTS), are characterised by a net negative charge and are therefore largely cell-impermeable. Considerable evidence indicates that their reduction occurs at the cell surface, or at the level of the plasma membrane via trans-plasma membrane electron transport. The implications of these new findings are discussed in terms of the use of tetrazolium dyes as indicators of cell metabolism and their applications in cell biology.
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MTT reduction reflects intracellular reductive activity rather than cell number per se, and may be influenced by mitochondrial function, redox state and metabolic stress independent of irreversible cell death.
“The underlying chemistry and cellular localization of tetrazolium dye reduction indicate that this signal reflects intracellular reductive activity rather than cell number per se, and may therefore be influenced by mitochondrial function, redox state and metabolic stress independent of irreversible cell death (Berridge et al., 2005).”
From Cytotoxicity across OECD methods is not a single endpoint: context-dependent roles, variability and implications for decision-making (2026) · cites this paper as Berridge et al., 2005
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Thoma, T., Löhr, L., Demuth, P., Hambruch, N., Oesch, F., & Landsiedel, R. (2026) · NAM Journal · doi:10.1016/j.namjnl.2026.100126
- Authors
- Berridge, M. V., Herst, P. M., & Tan, A. S.
- Affiliations
- Malaghan Institute of Medical Research (NZ)
- Corresponding author
- Berridge, Michael V.
- Venue
- Biotechnology Annual Review
- Publisher
- Elsevier
- Published
- 2005
- Volume
- 11
- Pages
- 127-152
- ISSN
- 1387-2656 (ISSN-L)
- OpenAlex
- W1849819493
- Cited by
- 2,222
- References
- 1
- Citations by year
- 2026: 115 · 2025: 101 · 2024: 138 · 2023: 131 · 2022: 127 · 2021: 130 · 2020: 137 · 2019: 171 · 2018: 172 · 2017: 162 · 2016: 179 · 2015: 167 · 2014: 126 · 2013: 111 · 2012: 46
- FWCI
- 166.38
- Type
- book chapter
- Language
- en
- Open access
- closed
- Locations
- Biotechnology annual review (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Toxicology › Bioactive Compounds and Antitumor Agents
- Physical Sciences › Chemistry › Organic Chemistry › Synthesis of Tetrazole Derivatives
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cancer and biochemical research
- Keywords
- tetrazolium dye, MTT, NBT, XTT, MTS, formazan, NADH, cell metabolic activity, cellular uptake, plasma membrane potential, cell viability assay
- MeSH terms
- Animals, Cell Division, Cell Survival, Cytological Techniques, Coloring Agents, Humans, Oxidation-Reduction, Tetrazolium Salts, Molecular Structure
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