Abstract
Structured illumination microscopy (SIM) provides wide-field optical sectioning in the focal plane by modulating the imaging information using fringe pattern illumination. For generating the fringe pattern illumination, a digital micro-mirror device (DMD) is commonly used due to its flexibility and fast refresh rate. However, the benefit of different pattern generation, for example, the two-beam interference mode and the three-beam interference mode, has not been clearly investigated. In this study, we systematically analyze the optical sectioning provided by the two-beam inference mode and the three-beam interference mode of DMD. The theoretical analysis and imaging results show that the two-beam interference mode is suitable for fast imaging of the superficial dynamic target due to reduced number of phase shifts needed to form the image, and the three-beam interference mode is ideal for imaging three-dimensional volume due to its superior optical sectioning by the improved modulation of the illumination patterns. These results, we believe, will provide better guidance for the use of DMD for SIM imaging and also for the choice of beam patterns in SIM application in the future.
Highlights
Fluorescence microscopy with optical sectioning provides unique benefits for the biological studies
We found the two-beam interference mode is suitable for fast imaging of the superficial dynamic target, whereas the three-beam interference mode is beneficial for imaging three-dimensional volume due to its superior optical sectioning capability
The relationships between the axial intensity distribution and the digital micro-mirror device (DMD) pattern periods for two-beam and three-beam interference modes are shown in Figure 3a,b, respectively
Summary
Fluorescence microscopy with optical sectioning provides unique benefits for the biological studies. Among fluorescence optical sectioning techniques, such as confocal laser scanning microscopy (CLSM) [1], two-photon microscopy (2PM) [2], and light-sheet fluorescence microscopy (LSFM) [3], structured illumination microscopy (SIM) is of growing interest due to its ability to integrate wide-field microscope and the high-speed. CLSM and 2PM, SIM provides higher-speed image acquisition because of its wide-field operating mode. The SIM has progressed through several advances over the years, with respect to exciting the specimen with high-contrast, fine illumination patterns for obtaining high-quality images. New opto-electro devices, such as the liquid crystal spatial light modulator (SLM) [4] and the digital micro-mirror device (DMD) which could be considered as a fast programmable grating [7], were applied to raise the switching speed of the fringe patterns. Compared to the SLM, which offers a refresh rate of hundreds of
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