Table 1.

Z′-factor calculated for varying cell density and luminescence read timepoint.

Cell Density	Z′-factor
	MNS-DMSO		TNF-DMSO		TNF-MNS
	45 min	60 min	45 min	60 min	45 min	60 min
1 × 104	0.41	0.43	−0.23	0.21	0.59	0.70
2 × 104	0.63	0.51	0.36	0.37	0.75	0.75
3 × 104	0.68	0.72	0.54	0.54	0.80	0.80
4 × 104	0.58	0.62	0.54	0.54	0.82	0.82

Z′ values calculated (Eq. 1) for different cell densities (1–4 × 10⁴ cells per 384-well) at either 45- or 60-minutes following dispensing of luciferin substrate. For column 1 (MNS-DMSO), MNS was used as the positive control and DMSO as the negative control. For column 2 (TNF-DMSO), TNF-α was used as the positive control and DMSO as the negative control. For column 3 (TNF-MNS), TNF-α was used as the positive control and MNS as the negative control. Overall, the LCA is most robust across all categories using a cell density of 3 × 10⁴.