eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2026-06-25 23 2 59304 article Korimelli Sridevi 1 Himaja Kommineni 1 Gunadeep Naidu Vennela 1 Department of Pharmacology, School of Pharmaceutical Sciences and Technologies, Jawaharlal Nehru Technological University, Kakinada, India In cancer research, drug development, and evaluation of treatment, the most critical factor is the accurate measurement of cell viability. In the last few decades, numerous approaches that focus on specific biochemical characteristics of the cells have been created to analyse the cytotoxicity, health, and proliferation of cells. Metabolic activity-based tetrazolium assays like MTT, XTT, and MTS are able to provide accurate and quantitative colorimetric assays based on the metabolic conversion of tetrazolium to formazan. Rapid and simple assessments based on the structural integrity of cell membranes and the differential staining of viable and dead cells are provided by Trypan Blue, Eosin, and Propidium Iodide.  Calcein-AM, Resazurin, and JC-1 are sensitive and reliable assays for the measurement of metabolic activity, evaluation of apoptosis, and assessment of the health of the mitochondria. This review provides a summary of the concepts, methods, advantages, and disadvantages of the described techniques. https://www.biotech-asia.org/vol23no2/comprehensive-overview-of-in-vitro-cell-viability-assays-in-cancer-research/ Alamar Blue; Calcein; Cell viability; MTT; Propidium iodide; Trypan Blue