Zhai Jiazhen, Jin Cheng, Kong Lingjie
State Key Laboratory of Precision Measurement Technology and Instruments, Department of Precision Instrument, Tsinghua University, Beijing 100084, China.
IDG/McGovern Institute for Brain Research, Tsinghua University, Beijing 100084, China.
J Phys Chem A. 2023 Mar 30;127(12):2873-2879. doi: 10.1021/acs.jpca.3c00325. Epub 2023 Mar 17.
Volumetric imaging of biodynamics at high spatiotemporal resolutions is vital in biomedical studies, in which Fourier light field microscopy (FLFM) is a promising technique. However, the commonly used wide-field illumination strategy in FLFM introduces intense out of depth-of-field background, which not only degrades the image quality, but also introduces reconstruction artifacts. Employing light sheet illumination is an effective way to alleviate the background and reduce photobleaching in light-field microscopy. Unfortunately, the introduction of light-sheet illumination often requires an extra objective and precise alignment, which increases the system complexity. Here, we propose the compact, hybrid light-sheet and FLFM (CLS-FLFM), which uses only a single objective to achieve both light-sheet illumination and Fourier light-field imaging simultaneously. With a micromirror under the objective, we focus the light sheet, which ensures selective-volume-illumination, on the imaging plane of the FLFM to perform volumetric imaging. We demonstrate the superior performance of CLS-FLFM in inhibiting background in both structural and dynamical imaging of larval zebrafish . We envision that CLS-FLFM finds wide applications in high-speed, background-inhibited volumetric imaging of biodynamics .
在生物医学研究中，以高时空分辨率对生物动力学进行体积成像至关重要，其中傅里叶光场显微镜（FLFM）是一项很有前景的技术。然而，FLFM中常用的宽场照明策略会引入强烈的离焦背景，这不仅会降低图像质量，还会引入重建伪影。采用光片照明是减轻光场显微镜背景并减少光漂白的有效方法。不幸的是，引入光片照明通常需要额外的物镜和精确对准，这增加了系统复杂性。在此，我们提出了紧凑的混合光片和FLFM（CLS-FLFM），它仅使用单个物镜即可同时实现光片照明和傅里叶光场成像。通过在物镜下方放置一个微镜，我们将确保选择性体积照明的光片聚焦在FLFM的成像平面上以进行体积成像。我们展示了CLS-FLFM在抑制斑马鱼幼体的结构和动态成像背景方面的卓越性能。我们设想CLS-FLFM将在生物动力学的高速、背景抑制体积成像中得到广泛应用。
