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Title: Multiplane Encoded Light-Sheet Microscopy for Enhanced 3D Imaging
Author: Zunino, Alessandro
Garzella, Francesco
Trianni, Alberta
Saggau, Peter
Bianchini, Paolo
Diaspro, Alberto
Duocastella, Martí
Keywords: Microscopis
Visualització tridimensional
Three-dimensional display systems
Issue Date: 3-Nov-2021
Publisher: American Chemical Society
Abstract: 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.
Note: Reproducció del document publicat a:
It is part of: ACS Photonics, 2021
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ISSN: 2330-4022
Appears in Collections:Articles publicats en revistes (Física Aplicada)

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