Development of low-coherence light sheet illumination microscope for fluorescence-free bioimaging

Zhiguang Xu; Timothy E. Holy

doi:10.1117/12.893569

9 September 2011 Development of low-coherence light sheet illumination microscope for fluorescence-free bioimaging

Zhiguang Xu, Timothy E. Holy

Proceedings Volume 8129, Novel Optical Systems Design and Optimization XIV; 812908 (2011) https://doi.org/10.1117/12.893569
Event: SPIE Optical Engineering + Applications, 2011, San Diego, California, United States

Abstract

Light Sheet Illumination Microscopy (LSIM) is an imaging modality featuring the novel arrangement with the illumination axis perpendicular to the detection axis. In this technology a well defined light sheet is generated and aligned precisely to the focal plane of the microscope objective and thus only the thin in-focus layer of the sample is illuminated and imaged, thereby avoiding out-of-focus light. Besides the inherent optical sectioning function, other advantages include fast imaging speed, high longitudinal resolution and decreased light-induced damage. Though promising, this microscopy is currently restricted to imaging fluorescently labeled tissue; in inspection of intact tissue using scattered light, the acquired images suffer from intense speckles because of the severe coherence in the illumination. This work aims to build a microscope capable of achieving intrinsic images of the fluorescence-free sample with reduced or eliminated speckles, by developing a low coherence light sheet illumination. To diminish the spatial coherence, the sample is illuminated with tens of independent sub-beams (without inter-coherence) illuminating the FOV (Field Of View) of the microscope with diverse incident angles. The temporal coherence is dramatically reduced by employing a supercontinuum laser with a broad spectrum as the light source. The new microscopy significantly extends the functionality of Light Sheet Illumination Microscopy and will enable many new bioimaging applications.

Citation Download Citation

Zhiguang Xu and Timothy E. Holy "Development of low-coherence light sheet illumination microscope for fluorescence-free bioimaging", Proc. SPIE 8129, Novel Optical Systems Design and Optimization XIV, 812908 (9 September 2011); https://doi.org/10.1117/12.893569

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