Abstract
Many biological structures of interest are beyond the diffraction limit of conventional microscopes and their visualization requires application of super-resolution techniques. Such techniques have found remarkable success in surpassing the diffraction limit to achieve sub-diffraction limited resolution; however, they are predominantly limited to fluorescent samples. Here, we introduce a non-fluorescent analogue to structured illumination microscopy, termed structured oblique illumination microscopy (SOIM), where we use simultaneous oblique illuminations of the sample to multiplex high spatial-frequency content into the frequency support of the system. We introduce a theoretical framework describing how to demodulate this multiplexed information to reconstruct an image with a spatial-frequency support exceeding that of the system’s classical diffraction limit. This approach allows enhanced-resolution imaging of non-fluorescent samples. Experimental confirmation of the approach is obtained in a reflection test target with moderate numerical aperture.
Highlights
Microscopy is critical in the biological sciences for its ability to visualize biological samples at the cellular level
Many biologically relevant structures are at sizes beyond the spatial frequency support of this diffraction limit and are physically unobservable using conventional optical techniques
We compared the resolution achieved between the orthogonally-illuminated (BF) and enhanced-resolution (SI) image of the Group 5 Element 4 set of bars of 22.1μm spacing
Summary
Microscopy is critical in the biological sciences for its ability to visualize biological samples at the cellular level. There are many subdivisions under this umbrella of general microscopy, and each are tailored towards specific visualization, design, and contrast requirements. Examples that have found widespread use include dark-field, phase-contrast, holographic, and fluorescent microscopies [1]. A critical factor that physically limits the optical resolution of microscopy in general is diffraction [2]. Many biologically relevant structures are at sizes beyond the spatial frequency support of this diffraction limit and are physically unobservable using conventional optical techniques. This has prompted many attempts to surpass this limit to achieve super-resolution
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