Table 1.

Acquisition parameters and default kinetic filtering criteria for iSiMREPS sensors for detecting miR-141 and EGFR exon 19 deletion mutant DNA in 10% v/v formamide.

iSMREPS sensor	Q₈C₆ QS₁₈CS₃ for detecting miR-141	Q₈C₆ QS₁₈CS₁₉ for detecting exon 19 deletion mutant DNA
Movie frames start-to-end	1–166	1–100
Exposure time per frame (s)	0.06	0.1
Acquisition time (s)	10	10
Intensity threshold per trace	200	500
S/N threshold per event	2	1.5
S/N threshold per trace	3.5	1.5
Minimum N_b+d	5	6
Maximum N_b+d	Inf	Inf
Minimum τ_{on, median} (s)	0.06	0.1
Maximum τ_{on, median} (s)	10	6
Minimum τ_{off, median} (s)	0.06	0.1
Maximum τ_{off, median} (s)	0.9	6
Maximum τ_{on, event} (s)	5	8
Maximum τ_{off, event} (s)	4	8

Note: τ_on and τ_off indicate target-bound (high-FRET) and non-target-bound (low-FRET) states, respectively. These criteria were obtained with the help of SiMREPS optimizer using several independent datasets with ≥ 10 FOVs both with and without target. Optimizer parameters were further tweaked manually to obtain a default that generally maximized true positives and minimized false positives. Each formamide variation experiment was individually optimized to allow each condition to be tested for its best possible performance.