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Lattice Selective Plane Illumination Microscopy (L-SPIM) for the analysis of subcellular dynamics in living specimens.

晶格选择性平面照明显微镜 (L-SPIM) 用于分析活体标本中的亚细胞动力学。

基本信息

批准号：
BB/T017899/1
负责人：
Steffen Scholpp
金额：
$ 83.31万
依托单位：
UNIVERSITY OF EXETER
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2020
资助国家：
英国
起止时间：
2020 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FT017899%2F1
关键词：
Lattice Selective Plane Illumination Microscopy

项目摘要

The discovery of the green fluorescent protein from the jellyfish Aequorea victoria has revolutionised our way in which we study cells in a tissue but also our approaches to investigate signalling events, organelle dynamics and interactions of organelles within the complex environment of the living cell. Parallel to the developments in GFP biology, there have been advances in fluorescence imaging methods and microscopical systems that make it possible to follow fluorescently labelled cells, organelles and cytoskeleton elements, to quantify their abundance and to probe their mobility and interactions. However, long-term imaging of complex 3D tissues with conventional laser-scanning microscopes is still one of the most significant obstacles in fluorescent microscopy as this microscopy induces phototoxicity and photobleaching. Selective plane illumination microscopes (SPIM) has been developed allowing long-term imaging by scanning the specimen with much less laser-induced damage such as phototoxicity and photobleaching. However, subcellular structures deep in tissue are still a challenge to resolve. Due to the recent development of Lattice SPIM, it is now possible to image small structures such as vesicles or microtubules over an extended scanning time deep in tissue without damaging the specimen. The combined advances in GFP biology and imaging methods are providing a massive opportunity for investigating the kinetic properties of organelles in living cells. The University of Exeter studies many aspects of biological and biomedical research, reaching from fungal-related plant disease research to signalling biology in vertebrate embryonic development. In the past, microscopical approaches such as confocal microscopy and electron microscopy have been used to investigate subcellular interactions. These observations can now be complemented and extended in real-time in living cells in complex organisms. The latest generation of Lattice SPIM is, therefore, a game-changer for cell biology - in bacteria, fungi, plants and animals. It is now possible to generate dynamic maps of organelles in living cells using a fluorescence microscope over a timescale of hours to days, which has not previously been possible. In this application, we seek support to purchase a modern Lattice SPIM, which will complement the existing facilities at the Exeter Bioimaging Centre.

来自维多利亚水母的绿色荧光蛋白的发现彻底改变了我们研究组织细胞的方式，也彻底改变了我们研究信号事件、细胞器动力学和复杂环境中细胞器相互作用的方法。活细胞。与GFP生物学的发展平行，荧光成像方法和显微镜系统也取得了进展，这使得可以跟踪荧光标记的细胞、细胞器和细胞骨架元素，量化它们的丰度并探测它们的流动性和相互作用。然而，复杂的三维组织与传统的激光扫描显微镜的长期成像仍然是荧光显微镜的最重要的障碍之一，因为这种显微镜诱导光毒性和光漂白。选择性平面照明显微镜（SPIM）已被开发，允许长期成像，通过扫描试样少得多的激光引起的损伤，如光毒性和光漂白。然而，组织深处的亚细胞结构仍然是一个有待解决的挑战。由于Lattice SPIM的最新发展，现在可以在延长的扫描时间内对组织深处的小结构（如囊泡或微管）进行成像，而不会损坏标本。GFP生物学和成像方法的综合进步为研究活细胞中细胞器的动力学特性提供了巨大的机会。埃克塞特大学研究生物学和生物医学研究的许多方面，从真菌相关的植物疾病研究到脊椎动物胚胎发育的信号生物学。在过去，显微镜的方法，如共聚焦显微镜和电子显微镜已被用来研究亚细胞相互作用。这些观察现在可以在复杂生物体的活细胞中实时补充和扩展。因此，最新一代的莱迪思SPIM将改变细菌、真菌、植物和动物细胞生物学的游戏规则。现在可以使用荧光显微镜在几小时到几天的时间尺度内生成活细胞中细胞器的动态图，这在以前是不可能的。在此申请中，我们寻求支持，以购买一台现代化的莱迪思SPIM，这将补充埃克塞特生物成像中心的现有设施。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Introduction: in vivo cell biology in zebrafish.

简介：斑马鱼体内细胞生物学。

DOI：
10.1007/s00418-020-01931-4
发表时间：
2020
期刊：
Histochemistry and cell biology
影响因子：
2.3
作者：
Scholpp S
通讯作者：
Scholpp S

The scaffolding protein Flot2 regulates cytoneme-based transport of Wnt3 in gastric cancer

DOI：
10.1101/2022.01.07.475396
发表时间：
2022-01
期刊：
bioRxiv
影响因子：
0
作者：
D. Routledge;Sally Rogers;H. Ashktorab;T. Phesse;S. Scholpp
通讯作者：
D. Routledge;Sally Rogers;H. Ashktorab;T. Phesse;S. Scholpp

Studying molecular interactions in the intact organism: fluorescence correlation spectroscopy in the living zebrafish embryo.

研究完整生物体中的分子相互作用：活斑马鱼胚胎中的荧光相关光谱。

DOI：
10.1007/s00418-020-01930-5
发表时间：
2020-11
期刊：
Histochemistry and cell biology
影响因子：
2.3
作者：
Dawes ML;Soeller C;Scholpp S
通讯作者：
Scholpp S

Wnt-7a-positive dendritic cytonemes induce synaptogenesis in cortical neurons

Wnt-7a 阳性树突状细胞因子诱导皮质神经元突触发生

DOI：
10.1101/2023.02.17.528927
发表时间：
2023
期刊：
影响因子：
0
作者：
Piers T
通讯作者：
Piers T

Cancer-associated fibroblasts influence Wnt/PCP signaling in gastric cancer cells by cytoneme-based dissemination of ROR2.

DOI：
10.1073/pnas.2217612120
发表时间：
2023-09-26
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Rogers S;Zhang C;Anagnostidis V;Liddle C;Fishel ML;Gielen F;Scholpp S
通讯作者：
Scholpp S