Structured Illumination Microscopy(SIM) is a suitable instrument for fluorescence imaging, especially dynamic imaging of live cells. It has two significant advantages of fast imaging speed and low excitation light energy density, and can reach the resolution of 100nm. In this article, we focus on the traditional reconstruction algorithm of SIM, which separates the spectrum with 9 raw images of 3-step phase-shifted picture in 3 orientations and operates the reconstruction with the correct displacement in Fourier space. However, without considering the initial phase error and the possibility that the displacement is a sub-pixel, there are artifacts in the results of the traditional reconstruction algorithm of SIM. To eliminate the artifacts and improve the imaging quality, we analyze the causes of various artifacts, and study the cross-correlation-based reconstruction algorithm of SIM, using the maximum cross-correlation value between the spectrum to get the correct displacement. Then, we simulate the illumination experimental parameters on the images of the reconstructed results. From the perspective of both hardware and software, we respectively consider the construction of the home-built SIM setup and the reconstruction software design, and finally realize the tri-color SIM system.
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