Abstract
Structured illumination microscopy (SIM) is a promising super-resolution technique for imaging subcellular structures and dynamics due to its compatibility with most commonly used fluorescent labeling methods. Structured illumination can be obtained by either laser interference or projection of fringe patterns. Here, we proposed a fringe projector composed of a compact multi-wavelength LEDs module and a digital micromirror device (DMD) which can be directly attached to most commercial inverted fluorescent microscopes and update it into a SIM system. The effects of the period and duty cycle of fringe patterns on the modulation depth of the structured light field were studied. With the optimized fringe pattern, [Formula: see text] resolution improvement could be obtained with high-end oil objectives. Multicolor imaging and dynamics of subcellular organelles in live cells were also demonstrated. Our method provides a low-cost solution for SIM setup to expand its wide range of applications to most research labs in the field of life science and medicine.
Highlights
Variations of Fluorescent microscopies are widely used as noncontact, minimally invasive imaging tools in molecular and cell biological studies
Structured illumination microscopy (SIM) is most attractive in live cell dynamic research due to reduction of photobleaching and photodamage, as well as its compatibility with most °uorescent labeling dyes and fast imaging speed.[10,11,12,13]
1:6Â spatial resolution improvement was obtained with our setup, benetted from the optimized fringe pattern
Summary
Variations of Fluorescent microscopies are widely used as noncontact, minimally invasive imaging tools in molecular and cell biological studies. The resolution of traditional °uorescent microscopy is. This is an Open Access article published by World Scientic Publishing Company. In the last 20 years, several di®erent kinds of super-resolution °uorescent microscopy techniques were developed, including Stimulated Emission Depletion microscopy (STED),[1,2,3] STochastic optical reconstruction microscopy (STORM),[4,5] Photo-activated localization microscopy (PALM),[6,7] and Structured illumination microscopy (SIM),[8,9] etc. SIM is most attractive in live cell dynamic research due to reduction of photobleaching and photodamage, as well as its compatibility with most °uorescent labeling dyes and fast imaging speed.[10,11,12,13]
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