Figure 1.

Schematic diagram of MEANS (a) and correlative images of a single cell acquired in TIRF (b) and MEANS (c) microscopy imaging modes. MEANS microscopy can be easily realized with a confocal microscope and takes advantage of axial interference between the incident and reflected electromagnetic field to generate an axially confined PSF ~100 nm above the reflective mirror surface. The merged image (d) of TIRF (green) and MEANS (red) shows that the MEANS approach complements the oblique illumination-based TIRF modality by optically sectioning a cell at a different axial layer which, in this case, is close to the mirror surface. The sample here is a Vero cell immunostained for tubulin using an AlexaFluor-488 secondary antibody. Scale bar=10 μm.