De novo endocytic clathrin coats develop curvature at early stages of their formation.

Sculpting a flat patch of membrane into an endocytic vesicle requires curvature generation on the cell surface, which is the primary function of the endocytosis machinery. Using super-resolved live cell fluorescence imaging, we demonstrate that curvature generation by individual clathrin-coated pits can be detected in real time within cultured cells and tissues of developing organisms. Our analyses demonstrate that the footprint of clathrin coats increases monotonically during the formation of pits at different levels of plasma membrane tension. These findings are only compatible with models that predict curvature generation at the early stages of endocytic clathrin pit formation. We also found that CALM adaptors associated with clathrin plaques form clusters, whereas AP2 distribution is more homogenous. Considering the curvature sensing and driving roles of CALM, we propose that CALM clusters may increase the strain on clathrin lattices locally, eventually giving rise to rupture and subsequent pit completion at the edges of plaques.

Design and optimization of microlens array based high resolution beam steering system.

High-resolution imaging and beam steering using 3 microlens arrays (MLA) is demonstrated. Small lateral displacement of one microlens array is sufficient for large angle beam steering. A prescan lens is added to the system to overcome the discrete addressing problem associated with microlens scanning systems. A hybrid method that uses both geometrical ray tracing optimization and physical optics simulation is introduced for the design and optimization of the MLA system. Feasibility of 1880 x 1880 resolution using f/2 aspherical MLAs and 752 x 752 resolution using f/5 spherical MLAs are demonstrated assuming 100 microm microlens pitch and 2mm clear aperture. The system is compact and suitable for endoscopic imaging and agile steering of large beams.

High-resolution beam steering using microlens arrays.

Imaging or beam-steering systems employing a periodic array of microlenses or micromirrors suffer from diffraction problems resulting from the destructive interference of the beam segments produced by the array. Simple formulas are derived for beam steering with segmented apertures that do not suffer from diffraction problems because of the introduction of a moving linear phase shifter such as a prescan lens before the periodic structure. The technique substantially increases the resolution of imaging systems that employ microlens arrays or micromirror arrays. Theoretical, numerical, and experimental results demonstrating the high-resolution imaging concept using microlens arrays are presented.