Table 1. Recommended acquisition parameters.

Parameter	Optimal
Well Shape*	U-bottom
Volume (Greiner U-bottom 96 well)**	>300 µL
Scan Pattern+	2×2
Flash Mode#	1 (400 Hz)
Number of Flashes	5

*Promotes hands-free centering of subjects.

**Filling the well to eliminate the meniscus effect (∼340 µL) produced the best results, however, this practice proved impractical for HTS.

⁺Promotes maximal throughput and consistently high Z'-factors (Table 2 & 3).

^#Lethality issues were evident at the 400 hertz level when volume was ≤100 µL.