Partial-frequency-spectrum reconstruction algorithm of SIM with reduced raw images

Abstract

In structured illumination microscopy (SIM), the imaging speed is expected to be increased to observe living cells. The conventional 2D SIM reconstruction algorithm (RA) requires nine raw images to reconstruct a super-resolution image. Here, we develop a partial-frequency-spectrum (PFS) reconstruction algorithm, based on the subtraction of frequency spectrum, which can reconstruct a super-resolution image by using six raw SIM images (two SIM images for each orientation). Our experiments of actin filament in bovine pulmonary artery endothelial (BPAE) cell imaging indicate that by the PFS algorithm, the frame rate increases. The PFS algorithm can resolve 120 nm in our experiment, which is equivalent to the reconstruction result of conventional 9-frame SIM. The PFS algorithm only requires the phase estimation of the three images. The reconstruction speed is about 5 times faster that of the conventional nine-images SIM method.