Figure 4
From: Highly efficient multicolor multifocus microscopy by optimal design of diffraction binary gratings
Optical characterization and validation of the fabricated 0.84 × π grating. (a) Simulations of the efficiency and uniformity of the diffraction through the grating as function of the wavelength. Overlaid are the measured values of the efficiency (blue squares) and uniformities (red circles). (b) Maximum intensity projection of a Z stack of fluorescent 200 nm diameter beads when imaged using multifocus microscope. (c) Validation of the plane spacing as measured by the beads. The spacing was found to be 470 nm for an emission centered at 620 nm in good agreement with the design value. (d) Zoom in on a point spread function of on the imaging planes, showing a diffraction limited PSF with no additional aberrations due to the grating motif. (e) Multifocus image of tubulin fibers in U2OS cells labeled with TMR molecules. (f) Multifocus image of the nuclear DNA labeled with Hoechst. The two images are taken with the same multifocus multicolor grating.
