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. 1991 Jan;63(1):119–125. doi: 10.1038/bjc.1991.24

Flow cytometric evaluation of hypoxic cells in solid experimental tumours using fluorescence immunodetection.

R J Hodgkiss 1, G Jones 1, A Long 1, J Parrick 1, K A Smith 1, M R Stratford 1, G D Wilson 1
PMCID: PMC1971656  PMID: 1989649

Abstract

Numerous methods have been proposed for the detection of hypoxic cells using nitroimidazoles labelled with both radioactive and stable isotopes where the isotopic label becomes bound as a result of reductive metabolism of the nitro group. A new probe for hypoxia, 7-(4'-(2-nitroimidazol-l-yl)-butyl)-theophylline, is described where an immunologically recognisable hapten (theophylline) is covalently linked to a 2-nitroimidazole. Bioreduction of the nitroimidazole leads to binding of bioreductive metabolites, and hence the theophylline side-chain, to intracellular molecules. Immunochemical procedures are then used to stain cells containing the bound theophylline using an FITC-conjugated anti-serum. Flow cytometric analysis of stained cells is facilitated by co-staining cellular DNA, which allows discrimination of single cells in the sample and rejection of cell clumps and debris. The alternative use of an immunoperoxidase-conjugated anti-serum has been used to demonstrate the localisation of hypoxic cells in frozen tumour sections.

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Selected References

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