Extended Data Fig. 6: Scatter plots of T-cell secretion-transition pattern over time.
From: Time-resolved assessment of single-cell protein secretion by sequencing
Shown were 6-color FACS analyses for the measurement of three cytokines simultaneously over two time periods, relating to Fig. 5. A total of four secretion windows (0, 1, 6, and 16 hour) with two consecutive secretion capture each were performed to track the secretion dynamics of single activated T cells. After the 1st-round of SCA, cells were stained in-situ with a first-set of fluorescent antibodies against IFN-γ, IL-2 and TNF-α to label cytokines secreted during this first period. After buffer exchange and supplementation with additional CapAbs, the cells were further incubated in the sealed nanowells for additional 45 minutes, and stained in situ with a second set of differently-labeled fluorescent antibodies to detect cytokines secreted during the second period.
