Advanced Multidimensional Optics to Investigate Biological Complexity at the Single Molecule Level in Living, Functional Cells
Advanced Multidimensional Optics to Investigate Biological Complexity at the Single Molecule Level in Living, Functional Cells
复制标题
先进的多维光学研究活体功能细胞中单分子水平的生物复杂性
DOI：
10.1016/j.bpj.2009.12.3192
复制
发表时间：
2010
影响因子：
3.4
通讯作者：
Harriman O
中科院分区：
文献类型：
--
作者：
Harriman O
荧光显微镜提供了一种微扰，非侵入性的手段来探测功能活细胞。该项目旨在开发能够同时成像多个生物分子（如基因组编码的荧光蛋白融合结构）的超分辨率、多色、多极化荧光显微镜。它将使动态调制的高对比度成像的带宽为100kHz，促进单细胞中的单个蛋白质分子的100毫秒跟踪到100nm的空间精度，并允许纳米级长度尺度的构象变化进行测量。它的构造将允许未来增加激光镊子，并将用于对单个活细胞进行研究。正在研究的是细胞过程，这些过程将单个蛋白质组装成功能性生物机器。显微镜将允许精确确定这些机器有多少组件，组装和拆卸过程的动力学，机械相互作用，分子稳定性以及不同生物过程的机器如何合作以在整个细胞中产生复合效应。
Fluorescence microscopy offers a minimally-perturbative, non-invasive means to probe functional living cells. This project aims to develop super-resolution, multi-colour, multi-polarization fluorescence microscopy capable of simultaneous imaging of multiple bio-molecules, such as genomically encoded fluorescent protein fusion constructs. It will enable dynamically-modulated high-contrast imaging to a bandwidth of∼ 1kHz, facilitate∼ millisecond tracking of single protein molecules in a single cell to∼ 10nm spatial precision, and permit nanoscopic length scale conformational change to be measured. It will be constructed to permit future addition of laser-tweezers and will be used to perform investigations on single living cells. Under investigation will be the cellular processes that bring about assembly of individual proteins into functional biological machines. The microscope will allow precise determination of how many components such machines have, dynamics of assembly and disassembly processes, mechanistic interactions, molecular stability and how machines of different biological processes may co-operate to produce compounded effects across the whole cell.