Confocal Raman microscopy is a useful tool to observe composition and constitution of label-free samples at high spatial resolution. However, accurate characterization of microstructure of tissue and its application in diagnostic imaging are challenging due to weak Raman scattering signal and complex chemical composition of tissue. We have developed a method to improve imaging speed, diffraction efficiency, and spectral resolution of confocal Raman microscopy.
View Article and Find Full Text PDFPoint scanning super-resolution microscopy techniques such as stimulated emission depletion (STED) microscopy are powerful tools to observe biological samples at sub-diffraction limited resolution in three dimensions. However, scanning the sample with only a single beam limits the imaging speed in these microscopes. Here, we propose a concept to increase this speed by introducing highly flexible multifocal illumination and detection.
View Article and Find Full Text PDFBiomolecular condensation is a widespread mechanism of cellular compartmentalization. Because the "survival of motor neuron protein" (SMN) is implicated in the formation of three different membraneless organelles (MLOs), we hypothesized that SMN promotes condensation. Unexpectedly, we found that SMN's globular tudor domain was sufficient for dimerization-induced condensation in vivo, whereas its two intrinsically disordered regions (IDRs) were not.
View Article and Find Full Text PDFUnderstanding cellular organization demands the best possible spatial resolution in all three dimensions. In fluorescence microscopy, this is achieved by 4Pi nanoscopy methods that combine the concepts of using two opposing objectives for optimal diffraction-limited 3D resolution with switching fluorescent molecules between bright and dark states to break the diffraction limit. However, optical aberrations have limited these nanoscopes to thin samples and prevented their application in thick specimens.
View Article and Find Full Text PDFOptical scatterfield imaging microscopy technique which has the capability of controlling scattered fields in the imaging mode is useful for quantitative nanoscale dimensional metrology that yields precise characterization of nanoscale features for semiconductor device manufacturing process control. To increase the sensitivity in the metrology using this method, it is required to optimize illumination and collection optics that enhance scatterfield signals from the nanoscale targets. Partial coherence of the optical imaging system is used not only for enhancing image quality in the traditional microscopy or lithography but also for increasing the sensitivity of the scatterfield imaging microscopy.
View Article and Find Full Text PDFWe propose a dual-detection confocal displacement sensor (DDCDS) with a compact fiber-based optical probe. This all-fiber-optic sensor probe is simple and robust, since it only requires simple alignment of a gradient refractive index lens and a double-clad fiber (DCF). The DDCDS is composed of two point detectors, one coupled to a single mode fiber and the other coupled to a multimode fiber, which are used to measure the light intensity from a core and an inner clad of a DCF, respectively.
View Article and Find Full Text PDFWe propose a new method for high-speed, three-dimensional (3-D) fluorescence imaging, which we refer to as dual-detection confocal fluorescence microscopy (DDCFM). In contrast to conventional beam-scanning confocal fluorescence microscopy, where the focal spot must be scanned either optically or mechanically over a sample volume to reconstruct a 3-D image, DDCFM can obtain the depth of a fluorescent emitter without depth scanning. DDCFM comprises two photodetectors, each with a pinhole of different size, in the confocal detection system.
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