弱光非线性结构光照明超分辨成像研究

项目名称： 弱光非线性结构光照明超分辨成像研究

项目编号： No.11504409

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 数理科学和化学

项目作者： 李思黾

作者单位： 中国科学院苏州生物医学工程技术研究所

项目金额： 24万元

中文摘要： 结构光照明荧光显微镜（SIM）是最具应用前景的超分辨成像设备之一，能够在可见光照明条件下，达到100nm横向分辨率，并且通过进一步引入非线性光学效应，开发非线性SIM技术，理论上可实现无限高分辨率完美成像。但是目前非线性SIM技术依赖于高功率照明光场，还没有合适的方法可以兼顾低光照功率、短光照时间、高成像分辨率，这极大地限制了非线性SIM技术在生物医学领域的应用。本项目研究弱光非线性结构光照明超分辨成像方法，创新性地提出低功率结构光激活―结构光激发的新型成像机制，并引入先进的光场调控方式以及使用新型高性能开关蛋白作为荧光探针，能够在弱光照明条件下(峰值功率比目前饱和非线性SIM技术低5个量级)，实现50nm横向分辨率快速超分辨显微成像。本项目的实施可在超分辨成像基础理论研究中取得原始创新成果，并为生物学和医学前沿课题研究提供先进的实验技术。

中文关键词： 超分辨荧光显微成像；结构光照明显微成像；光场调控；非线性光学效应

英文摘要： Structured-illumination microscopy (SIM) is one of the most promising super-resolution imaging devices in life sciences. Previously, linear SIM has achieved ~100nm resolution under visible light-wave illumination with greater resolution extension possible if appropriate nonlinear fluorescence phenomena are introduced, then an implementation of nonlinear structured-illumination microscopy (NL-SIM) can be generated that is of theoretically unlimited resolution. Unfortunately, in their most basic form (i.e., under conditions of saturated nonlinearity), NL-SIM methods require extreme peak intensities from the incident laser―on the order of MW/cm^2 , and to our knowledge there have been no published results of fast super resolution NL-SIM methods under ultralow light intensity with ultra-short exposure time. This project proposes a novel method to implement weak-light nonlinear structured-illumination microscopy (WNL-SIM). By exploring unique super-resolution imaging mechanism with both activating fields and exciting fields are sinusoidal structured, as well as new approaches to generate maximal contrast optical structured fields, it will reveal bio-samples at 50nm resolution using light intensities on the order of 10W/cm^2 that is five orders of magnitude lower than those needed for saturation NL-SIM. This project will achieve original innovation in basic bio-imaging research and a biologically compatible super-resolution imaging method for biology and fundamental medicine study.

英文关键词： superresolution microscopy;Structured-illumination microscopy;light field manipulation;Nonlinear optical effect