Recent advances in fluorescence nanoscopy have pushed resolution to the 1–10 nm range,enabling the direct visualization of individual molecules even in crowded biological environments.Achieving this level of precisio...Recent advances in fluorescence nanoscopy have pushed resolution to the 1–10 nm range,enabling the direct visualization of individual molecules even in crowded biological environments.Achieving this level of precision requires rigorous sample drift control.Techniques such as MINFLUX and RASTMIN,which rely on keeping the sample fixed within an excitation pattern,demand active drift correction to achieve their theoretical nanometer-scale resolution limits.Here,we present an active stabilization system for super-resolution microscopy that delivers sub-nm precision for hours.Featuring a simple optical design,the system can be added as a separate module to any fluorescence microscope.We also provide an open-source control software including a user-friendly graphical interface readily adaptable to different setups.We demonstrate the adaptability and performance of the stabilization system with p-MINFLUX and RASTMIN measurements performed in two different setups,reaching the theoretical Cramér-Rao Bound and resolving~10 nm distances within DNA origami structures.展开更多
基金funded by Consejo Nacional de Investigaciones Científicas y Técnicas(CONICET)Swiss National Science Foundation project 200021_184687(G.P.A.)+1 种基金Agencia Nacional de Promoción Científica y Tecnológica(ANPCYT),projects PICT-2017-0870(F.D.S.),and PICT-2021-01216(F.D.S.)funded by Warsaw University of Technology within the Excellence Initiative:Research University(IDUB)programme.
文摘Recent advances in fluorescence nanoscopy have pushed resolution to the 1–10 nm range,enabling the direct visualization of individual molecules even in crowded biological environments.Achieving this level of precision requires rigorous sample drift control.Techniques such as MINFLUX and RASTMIN,which rely on keeping the sample fixed within an excitation pattern,demand active drift correction to achieve their theoretical nanometer-scale resolution limits.Here,we present an active stabilization system for super-resolution microscopy that delivers sub-nm precision for hours.Featuring a simple optical design,the system can be added as a separate module to any fluorescence microscope.We also provide an open-source control software including a user-friendly graphical interface readily adaptable to different setups.We demonstrate the adaptability and performance of the stabilization system with p-MINFLUX and RASTMIN measurements performed in two different setups,reaching the theoretical Cramér-Rao Bound and resolving~10 nm distances within DNA origami structures.
