Table 1. List of cytotoxicity assays.

Cytotoxicity assays, some of which were used in this study, listed along with the brief description of their key features.

Assay/dye	Type(s) of cell death detected	Necessary equipment	Key features
MTT, CKK-8, alamarBlue (resazurin)	Apoptosis/Necrosis	Spectrophotometer	Inexpensive, rapid; endpoint assay; dependent on enzymes’ activity (exclusively mitochondrial in case of MTT) and does not discriminate between modes of cell death1,10
LDH release	Necrosis	Spectrophotometer	Rapid, independent of mitochondrial enzymes’ activity; expensive for high-throughput tests; detects necrotic cells with compromised plasma membrane11,12
Trypan blue (TB)	Apoptosis/Necrosis	Microscope	Cell-impermeant; does not discriminate between modes of cell death; laborious and not suitable for high-throughput screening; more difficult to use with adherent cells; prone to subjective judgment of the user, but is considered the standard cell viability measurement method13
Acridine orange (AO)	Apoptosis/Necrosis/Necroptosis	Fluorescence microscope	A nucleic acid dye with unique spectral properties, can distinguish between apoptosis and necrosis/necroptosis14
Hoechst 33342, DAPI	Apoptosis	Fluorescence microscope or flow cytometer	Cell-permeable; inappropriate on its own to monitor cell death; useful for co-staining; can be used to assess chromatin condensation and nuclei fragmentation in early apoptosis; can be paired with propidium iodide to distinguish apoptosis from necrosis15,16
Propidium iodide (PI)	Late apoptosis/Necrosis	Fluorescence microscope or flow cytometer	Cell-impermeant intercalator; detects both late apoptosis and necrosis modes of cell death17. Toxic and permeable after long incubation times18