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结构光照相和荧光显微镜在生物成像中的应用。

Structured illumination phase and fluorescence microscopy for bioimaging.

出版信息

Appl Opt. 2023 Jun 20;62(18):4871-4879. doi: 10.1364/AO.486718.

Abstract

This study presents a dual-modality microscopic imaging approach that combines quantitative phase microscopy and fluorescence microscopy based on structured illumination (SI) to provide structural and functional information for the same sample. As the first imaging modality, structured illumination digital holographic microscopy (SI-DHM) is implemented along the transmission beam path. SI-DHM acts as a label-free, noninvasive approach and provides high-contrast and quantitative phase images utilizing the refractive index contrast of the inner structures of samples against the background. As the second imaging modality, structured illumination (fluorescence) microscopy (SIM) is constructed along the reflection beam path. SIM utilizes fluorescent labeling and provides super-resolution images for specific functional structures of samples. We first experimentally demonstrated phase imaging of SI-DHM on rice leaves and fluorescence (SIM) imaging on mouse kidney sections. Then, we demonstrated dual-modality imaging of biological samples, using DHM to acquire the overall cell morphology and SIM to obtain specific functional structures. These results prove that the proposed technique is of great importance in biomedical studies, such as providing insight into cell physiology by visualizing and quantifying subcellular structures.

摘要

本研究提出了一种结合定量相显微镜和基于结构光照相(SI)的荧光显微镜的双模式微观成像方法,为同一样品提供结构和功能信息。作为第一种成像模式,结构光照相数字全息显微镜(SI-DHM)沿透射光束路径实现。SI-DHM 作为一种无标记、非侵入式方法,利用样品内部结构与背景之间的折射率对比提供高对比度和定量相位图像。作为第二种成像模式,结构光照相(荧光)显微镜(SIM)沿反射光束路径构建。SIM 利用荧光标记,为样品的特定功能结构提供超分辨率图像。我们首先在水稻叶片上实验演示了 SI-DHM 的相位成像,以及在小鼠肾脏切片上演示了 SIM 的荧光成像。然后,我们演示了生物样品的双模式成像,使用 DHM 获得整体细胞形态,使用 SIM 获得特定功能结构。这些结果证明,该技术在生物医学研究中具有重要意义,例如通过可视化和量化亚细胞结构来洞察细胞生理学。

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