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通过双光子激发荧光(TPEF)、二次谐波产生(SHG)和三次谐波产生(THG)显微镜联合技术对线虫细胞结构进行体内成像。

In vivo imaging of cellular structures in Caenorhabditis elegans by combined TPEF, SHG and THG microscopy.

作者信息

Gualda E J, Filippidis G, Voglis G, Mari M, Fotakis C, Tavernarakis N

机构信息

Institute of Electronic Structure and Laser, Foundation of Research and Technology-Hellas, 71110, Heraklion, Crete, Greece.

出版信息

J Microsc. 2008 Jan;229(Pt 1):141-50. doi: 10.1111/j.1365-2818.2007.01876.x.

Abstract

In this study, we use combined two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG) and third-harmonic generation (THG) measurements to image cellular structures of the nematode Caenorhabditis elegans, in vivo. To our knowledge, this is the first time that a THG modality is employed to image live C. elegans specimens. Femtosecond laser pulses (1028 nm) were utilized for excitation. Detailed and specific structural and anatomical features can be visualized, by recording THG signals. Thus, the combination of three image-contrast modes (TPEF-SHG-THG) in a single instrument has the potential to provide unique and complementary information about the structure and function of tissues and individual cells of live biological specimens.

摘要

在本研究中,我们使用双光子激发荧光(TPEF)、二次谐波产生(SHG)和三次谐波产生(THG)相结合的测量方法对秀丽隐杆线虫的细胞结构进行活体成像。据我们所知,这是首次使用THG模态对活的秀丽隐杆线虫标本进行成像。使用飞秒激光脉冲(1028纳米)进行激发。通过记录THG信号,可以可视化详细且特定的结构和解剖特征。因此,单一仪器中三种图像对比度模式(TPEF-SHG-THG)的组合有潜力提供关于活生物标本的组织和单个细胞的结构与功能的独特且互补的信息。

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