基于三通道的超分辨率显微镜成像

Super Resolution Microscope Imaging Based on Three Channels

显微技术经过快速发展,已经突破了光学衍射极限,目前主要包含受激发射损耗显微术(STED)、结构光照明显微镜(SIM)、光激活定位显微成像(PALM)、随机光学重构显微术(STORM)、基于最少光子数的纳米尺度定位(MINFLUX)、结合结构光照明技术的MINFLUX技术变体(SIMFLUX)等技术。STORM技术具有优越性,在其基础上叠加多色成像技术(目前有6种),本文介绍了目前最新的多色成像技术以及分光成像实现的三通道成像技术。分光成像实现的三通道成像存在光谱串色、通道对齐误差等影响,基于此介绍了相关的优化算法原理。展示了在三通道STORM显微成像平台上实现的COS-7细胞成像。说明三通道STORM显微成像的优越性。

Microscopy technology has undergone rapid development and has surpassed the limits of optical diffraction.Currently,it mainly includes stimulated emission depletion microscopy(STED),structured illumination microscopy(SIM),photoactivation localization microscopy(PALM),stochastic optical reconstruction microscopy(STORM),minimal photon fluxes(MINFLUX) based on the localization of the minimum number of photons,and a variant of MINFLUX technology combining structured illumination microscopy(SIMFLUX).STORM technology has superiority,and on top of it,there are six multi-color imaging technologies currently available.This article introduces the latest multi-color imaging technology and a three-channel imaging technique implemented using spectral imaging.However,three-channel imaging using spectral imaging has issues such as spectral crosstalk and channel alignment errors.Therefore,relevant optimization algorithm principles are introduced.The article also demonstrates the imaging of COS-7 cells on a three-channel STORM microscopy platform,showcasing the superiority of three-channel STORM microscopy.