Table 1.

Advantages and disadvantages of methods used for drug cytotoxicity analysis.

Method	Advantages	Disadvantages	Application
MTT, MTS, XTT, WST-1	Inexpensive; Highly accessible; Relatively fast; Simple in execution.	Does not detect dead cells; Cells are analyzed in bulk; Additional steps in protocol may increase data variability (in case of MTT); Toxic to cells.	High-throughput screening
LIVE/DEAD assay	Allows to evaluate numbers of both viable and dead cells; Rapid process; Simple in execution; Moderately accessible; Can be optimized for microscopic studies.	Expensive; Cells are analyzed in bulk; Requires optimization for each cell line; Limited sensitivity in adherent cells [52].	Screening of small compound collections
Annexin V flow cytometry analysis	Estimates the fractions of viable cells, cells in early apoptosis, and cells in late apoptosis/necrosis; Analyzes individual cells.	Provides no information on total cell number; Does not discern between cells in late apoptosis and necrosis. Analysis of adherent cells requires additional procedures that may affect the viability; Cannot be scaled for high-throughput analysis.	Study of apoptosis in a limited number of samples; Identification of drug-sensitive and drug-resistant subpopulations.
Western blotting	Allows to estimate molecular mechanisms of drug action; High sensitivity.	Provides no information on cell numbers; Cells are analyzed in bulk; Requires high-quality antibodies against cell death markers; Cannot be scaled for high-throughput analysis.	Study of molecular mechanisms of drug action in a small number of samples.