《Super-resolution Microscopy》细胞成像技术小组展示.pptxVIP

Super-resolution Microscopy ClassificationNear field : NSOMFar field :PALM/STORM/SOFI STED/GSD SIM01Near field PART ONE NSOMNSOM (Near-Field scanning optical microscopy)Eric Berzig(N.E. Dickenson et al.,Surface Analysis and Techniques in Biology, 2014.)Application：Biological studies(CH Ke et al. , BBA-BIOMEMBRANES, 2015.)Problems Low working distance with the sample Limited to study surface Long scan times for large sample areasFar field 02Single molecule fluorescence position PALM/STORM/SOFIPoint spread function STED/GSDStructured illumination SIM PART TWO PALM (Photo-activation localization microscopy)(Betzig E. et al , Science, 2006. )STORM (Stochastic optical reconstruction microscopy)red laser (633 nm) excite fluorescence from Cy5 and to switch Cy5 to dark state. green laser (532 nm) return Cy5 to luorescent state.(Michael J Rust et al.,Nature Methods , 2006.)Applications: Supermolecular polymers and organelles STORM(KeXu et al., Science, 2013. )Interactions between biological macromolecules PALMpositivecontrolnegativecontrol(Eilon Sherman et al., Methods, 2013.) Single molecule tracking PALM Quantitative analysis of the subcellular level PALMProblems Long scan timesSTORM:Can not measure the living cellsCan not quantitativePALM:Only observe exogenous proteins expressed Too much background noise SOFI (Super-Resolution Optical Fluctuation Imaging)（Xi Zhang et al., ACS Nano, 2015.)（Dertinger.T et al., PNAS, 2009.）Applications：small moleculars and organelles(Dertinger. T. et al., Quarterly reviews of biophysics, 2013.) 3D-imaging(Geissbuehler.S. et al.,Nature communications , 2014.)Problems Much requirements for light-emitting molecule Not enough high resolution STED (Stimulated emission depletion ) Microscopy(Hell S.W. et al., Optical Society of America, 1994.) (Willig KI et al ., Nature, 2006.)Applications: Smaller molecular (Willig KI et al ., Nature, 2006.)Living cell (Natalia M. Mishina et al . Nano, 2015.)Living slices (Ronan Chéreau et al ., Met