Mesoscopic axially swept oblique plane microscope for the imaging of freely moving organisms with near-isotropic resolution.

IF 2.9 2区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Biomedical optics express Pub Date : 2024-11-06 eCollection Date: 2024-12-01 DOI:10.1364/BOE.537262

Samuel Davis, Jon-Richard Sommernes, Sebastian Hambura, Levin Riedel, Alejandro Gil, Aissam Ikmi, Florian Ströhl, Robert Prevedel

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引用次数: 0

Abstract

Rapid three-dimensional imaging over extended fields of view (FOVs) is crucial to the study of organism-wide systems and biological processes in vivo. Selective-plane illumination microscopy (SPIM) is a powerful method for high spatio-temporal resolution in toto imaging of such biological specimens. However, typical SPIM implementations preclude conventional sample mounting and have anisotropic imaging performance, in particular when designed for large FOVs over 1 mm diameter. Here, we introduce axial sweeping of the illumination into a non-orthogonal dual-objective oblique plane microscope (OPM) design, thereby enabling the observation of freely moving animals over millimeter-sized FOVs, at close to isotropic, sub-cellular resolution. We apply our mesoscopic axially swept OPM (MASOPM) to image the behavioral dynamics of the sea anemone Nematostella vectensis over 1 × 0.7 × 0.4 mm at 1.7 × 2.6 × 3.7 µm resolution and 0.5 Hz volume rate.

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用于自由移动生物成像的介观轴向扫描斜面显微镜，分辨率接近各向同性。

扩展视场（FOV）的快速三维成像对于研究整个生物系统和体内生物过程至关重要。选择性平面照明显微镜（SPIM）是对此类生物样本进行高时空分辨率整体成像的有力方法。然而，典型的 SPIM 实现方法不包括传统的样品安装，而且成像性能各向异性，尤其是在为直径超过 1 毫米的大视野设计时。在这里，我们在非正交双目标斜面显微镜（OPM）的设计中引入了轴向扫描照明，从而能够在毫米大小的视场中观察自由移动的动物，分辨率接近各向同性的亚细胞分辨率。我们应用我们的介观轴向扫描 OPM（MASOPM），以 1.7 × 2.6 × 3.7 µm 的分辨率和 0.5 Hz 的体积率，对海葵 Nematostella vectensis 在 1 × 0.7 × 0.4 mm 上的行为动态进行成像。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biomedical optics express BIOCHEMICAL RESEARCH METHODS-OPTICS

CiteScore

6.80

自引率

11.80%

发文量

633

审稿时长

1 months

期刊介绍： The journal''s scope encompasses fundamental research, technology development, biomedical studies and clinical applications. BOEx focuses on the leading edge topics in the field, including: Tissue optics and spectroscopy Novel microscopies Optical coherence tomography Diffuse and fluorescence tomography Photoacoustic and multimodal imaging Molecular imaging and therapies Nanophotonic biosensing Optical biophysics/photobiology Microfluidic optical devices Vision research.