Table 1:

Example SUIT protocol to assess mitochondrial membrane potential in freshly isolated T-cells and monocytes using the 0.5 mL chambers

SUIT labels	Injection	Injection volume (μL)	Final concentration in 0.5mL chamber	Comments/Expected observations
TMRM calibration	0.05 mM TMRM	2.5 X 4	0.25–1.0 μM	Increase in fluorescence per injection
Pre-Cell	TMRM reading just prior to cell addition
Cell	Cells suspended in Mir05	20	Mono: 5 million / mL	Read for 10 minutes. Decline in fluorescence as TMRM is taken in.
			T-cell: 10 million / mL
DIG	1 mg/mL digitonin	2	4 μL/mL	Read for 20 minutes. Stabilization of fluorescence.
SUCC	1 M succinate	2.5	5 mM	Sharp decline in TMRM fluorescence (increased mMP)
MPG	0.1 M malate	5	1.0 mM	Simultaneous injections. Small decline in TMRM fluorescence (increased mMP)
	1.0 M glutamate	5	10 mM
	0.5 M pyruvate	5	5 mM
D1-D10	ADP (0.5–500mM)	titrat.	0.01– 4.0 mM	Notice ADP-driven rise in TMRM fluorescence (decrease in mMP)
OMY	1 mg/mL oligomycin	1	2 μg/ml	Small decrease in fluorescence (increase in mMP)
FCCP	0.25 mM FCCP	1 X 3	0.5–1.5 μM	Sharp increase in fluorescence (depletion of mMP)
ROT	0.25 mM Rotenone	1	0.5 μM	Decline in respiration. mMP values are not relevant after FCCP addition.
AMA	1.25 mM Antimycin A	1	2.5 μM	Inhibition of mitochondrial respiration