Electro-optics and Microelectronics Seminar
Speaker: Doron Shterman
Affiliation: Dept. of Electrical Engineering, Technion
Multiple Moire’ Structured Illumination patterns for spatial resolution enhancement in florescence microscopy
Biological research nowadays is heavily dependent on the capabilities of modern microscopy, especially on their spatial and temporal resolution. Spatial resolution in conventional light microscopy is however limited by diffraction, as formulated by Ernst Abbe in 1873. In an attempt to overcome this limitation several super-resolution techniques have evolved in the past decades, based on fluorescence microscopy. Methods like Stimulated Emission Depletion and Photo-activated Localization Microscopy, can achieve sub 50nm resolution yet requires relatively long acquisition times and\or rely damaging intensities, making them inadequate for dynamic in vivo studies. Structured Illumination Microscopy (SIM), on the other hand, is a wide-field super-resolution technique with superior temporal capabilities, while its spatial resolution is somewhat inferior reaching conventionally roughly 100nm.
In this work we proposed and demonstrated, theoretically and empirically, a novel super-resolution microscopy concept based on combination of structured illumination and total internal reflection techniques. Utilizing multiple Moire’ illumination patterns, precisely and independently controlled by spatial light modulator (SLM), and high index material, sub 50nm imaging can be achieved while preserving SIM temporal resolution capability.
As a proof of concept, we demonstrated resolution enhancement of more than 3 times beyond the diffraction limit, utilizing combination of low and high numerical aperture objective lenses as a substitute for the high index material.
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