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2021-11-03

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Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/183314

Multiplane Encoded Light-Sheet Microscopy for Enhanced 3D Imaging

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https://doi.org/10.1021/acsphotonics.1c01401

Resum

Light-sheet microscopes have become the tool of choice for volumetric imaging of large samples. Based on a wide-field acquisition scheme, they are capable of optical sectioning at diffraction-limited resolution and minimal overall photodamage. Unfortunately, traditional architectures are limited in speed because 3D images are collected by either sample translation or synchronized movement of both light-sheet and detection objective lens. A promising solution avoiding slow mechanical movements is to extend the depth-of-field of the microscope and moving only the light-sheet. However, this normally comes at the cost of losing light and contrast, compromising the signal-to-noise ratio of the images. Here, we propose an innovative technique devoted to restoring the quality of the images, while preserving the speed of extended depth-of-field microscopes. It is based on generating a stack of parallel light-sheets using a pair of orthogonal acousto-optic deflectors, enabling the simultaneous illumination of different sample planes. Given the extended depth-of-field, all such planes appear in focus and can be acquired in a superimposed single frame. By applying a single-step inversion algorithm, we can decode a stack of frames into a volumetric image whose signal-to-noise ratio and contrast are greatly enhanced. We provide a detailed theoretical framework of the method and demonstrate its feasibility with volumetric images of kidney cell spheroids.

Matèries

Microscopis, Visualització tridimensional

Matèries (anglès)

Microscopes, Three-dimensional display systems

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Citació

ZUNINO, Alessandro, et al. Multiplane Encoded Light-Sheet Microscopy for Enhanced 3D Imaging. ACS Photonics. 2021. ISSN 2330-4022. [consulta: 11 de maig de 2026]. Disponible a: https://hdl.handle.net/2445/183314

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