title: Assessment of Cell Viability in Intact Glandular Tissue in Chironomus ramosus using Dye-exclusion and Colorimetric Assays.
abstract: Conventionally, dye-exclusion test for determining cell viability has been restricted only for cells in suspension in tissue culture. In this paper, salivary gland of Chironomus has been proposed as a simple tissue model system where dye-exclusion test can be reliably employed for the intact gland. We have compared suitability of commonly used vital dyes and nigrosin was found suitable for the salivary gland cells. Biochemical tests using tetrazolium salts are also commonly used for determining quantitative indices of cell viability in metabolically active cells. Ours is the first attempt to extend the same technique for the whole tissue. We standardized the conditions and prepared a protocol for MTT-based colorimetric assay suitable for the salivary gland of Chironomus. A strong correlation (r(2) = 0.9893) was obtained where increasing O.D. correlated linearly with the number of live glands. We concluded that nigrosin dye-exclusion and MTT metabolic inclusion assays are suitable methods for the viability test of metabolically active intact salivary gland of Chironomus which can serve as a potential model for the assessment of cytotoxicity in future.
authors.affiliation: Department of Zoology, University of Pune, Pune, 411007, India, bbnath@unipune.ernet.in.
authors.forename: B B
authors.initials: BB
authors.lastname: Nath
authors.forename: A A
authors.initials: AA
authors.lastname: Babrekar
authors.forename: B
authors.initials: B
authors.lastname: Parthasarathy