title: Flow cytometric evaluation of hypoxic cells in solid experimental tumours using fluorescence immunodetection.
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.
authors.affiliation: Gray Laboratory of the Cancer Research Campaign, Mount Vernon Hospital, Northwood, Middlesex, UK.
authors.forename: R J
authors.initials: RJ
authors.lastname: Hodgkiss
authors.forename: G
authors.initials: G
authors.lastname: Jones
authors.forename: A
authors.initials: A
authors.lastname: Long
authors.forename: J
authors.initials: J
authors.lastname: Parrick
authors.forename: K A
authors.initials: KA
authors.lastname: Smith
authors.forename: M R
authors.initials: MR
authors.lastname: Stratford
authors.forename: G D
authors.initials: GD
authors.lastname: Wilson
mesh_terms: D000818:Animals; D015687:Cell Hypoxia / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D009374:Neoplasms, Experimental / Q000473:pathology*; D009593:Nitroimidazoles / Q000378:metabolism; D012513:Sarcoma, Experimental / Q000473:pathology*; D013806:Theophylline / Q000378:metabolism
chemical_list: D009593:Nitroimidazoles; D013806:Theophylline