Abstract
Far-field fluorescence microscopy has made great progress in the spatial resolution, limited by light diffraction, since the super-resolution imaging technology appeared. And stimulated emission depletion (STED) microscopy and structured illumination microscopy (SIM) can be grouped into one class of the super-resolution imaging technology, which use pattern illumination strategy to circumvent the diffraction limit. We simulated the images of the beads of SIM imaging, the intensity distribution of STED excitation light and depletion light in order to observe effects of the polarized light on imaging quality. Compared to fixed linear polarization, circularly polarized light is more suitable for SIM on reconstructed image. And right-handed circular polarization (CP) light is more appropriate for both the excitation and depletion light in STED system. Therefore the right-handed CP light would be the best candidate when the SIM and STED are combined into one microscope. Good understanding of the polarization will provide a reference for the patterned illumination experiment to achieve better resolution and better image quality.
Highlights
Fluorescence microscopy is widely used in lifescienceeld
To demonstrate the e®ects of light polarization on structured illumination microscopy (SIM) imaging, we simulated two-beam interference structured illumination with di®erent polarization
We found that thexed linear polarization (LP) of tworst-order beams in pattern orientation 90 has the lowest modulation contrast
Summary
The spatial resolution of conventional °uorescence microscopy is limited because of the di®raction of light, which is called the Abbe di®raction limit.[1] Over the past decade the researchers have developed several super resolution techniques, such as stimulated emission depletion (STED). This is an Open Access article published by World Scientic Publishing Company. Microscopy,[2,3] structured illumination microscopy (SIM),[4,5] photoactivated localization microscopy (PALM).[6] and stochastic optical reconstruction microscopy (STORM)[7] to circumvent the di®raction limit These super resolution techniques can be divided into two categories. To demonstrate the in°uence of polarized light of di®erent modes on the quality of images of SIM and STED, we simulated images of °uorescence beads for SIM and scanning spot of the excitation light and depletion light with polarization for STED
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