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TIRF example

Total internal reflection fluorescence (TIRF) microscopy example images of a NIH3T3 cell transfected with a tubulin-GFP plasmid construct. A) Example cell under normal GFP epifluorescence, B) the same cell when viewed with TIRF microscopy with a 140 nm thickness setting, C) the same cell with a TIRF thickness setting of 70 nm.
NB: Narrowing the TIRF thickness setting in (C) reduces the number of illuminated GFP labeled microtubules as now only those microtubules directly adjacent to the coverslip are getting fluorescently excited. Images were collected with a Leica DMI 6000B TIRF microscope.
Image taken from Francis RJ et al. Connexin43 modulates cell polarity and directional cell migration by regulating microtubule Dynamics. unpublished manuscript.