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. 2016 Aug 12;7:12471. doi: 10.1038/ncomms12471

Figure 4. SRRF used for super-resolution live-imaging of Jurkat T cells transiently expressing LifeAct-GFP.

Figure 4

(a) Conventional TIRF microscopy (TIRF) versus SRRF images (SRRF) of Jurkat T cells transfected with LifeAct-GFP dropped on an anti-CD3-coated coverslip and imaged for 180 s at 1 super-resolution f.p.s. (b) Conventional TIRF and SRRF images of Jurkat T cells expressing LifeAct-GFP imaged on coverslips coated with anti-CD28 alone, anti-CD3 alone or in combination (anti-CD3 and -CD28) to stimulate an immunological synapse formation (highlighted area corresponds to the region used for PIV analysis). (c) PIV analysis of corresponding Supplementary Movies 6–8 shows notable retrograde actin flow in anti-CD3 but not in anti-CD28 stimulated Jurkat T cells. A colour-coded measure of flow directionality and speed is plotted for the blue highlighted regions. White arrows in colour wheel are representative of flow direction, central colour (minimum intensity) corresponds to 0 μm min−1, saturated colours (maximum intensity) correspond to 38.4 μm min−1. Scale bars, 5 μm.