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. 2019 Jul 3;27(14):19950–19972. doi: 10.1364/OE.27.019950

Fig. 3.

Fig. 3

vaTIRF imaging of RhoA activity using a FRET biosensor. Mouse Embryonic Fibroblast (MEF) cell stably expressing a RhoA biosensor [54] imaged sequentially with epifluorescence illumination (a), fixed angle TIRF (b) and vaTIRF (c) to compare different imaging modes. FRET/Donor ratio images from epifluorescence and vaTIRF showed RhoA activity in cell protrusions (outward pointing arrow) and retractions (inward pointing arrow), while uneven illumination with fixed angle TIRF produced spurious activation maps (e.g. inward pointing arrow head).