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Abstract
En 中文
Spatial resolution of conventional far-field fluorescence microscopy is limited by diffraction of light. Single-molecule localization microscopy (SMLM), such as (direct) stochastic optical reconstruction microscopy (dSTORM/STORM), and (fluorescence) photoactivation localization microscopy (fPALM/PALM), can break this barrier by localizing single emitters and reconstructing super-resolution image with much higher precision. Nevertheless, a SMLM measurement needs to record a large number of image frames and takes considerable recording time. In this process, sample drift becomes a critical problem and cannot be neglected. In this Letter, we present a sub-nanometer precision, low-cost sample drift correction method based on minimizing normalized root-mean-square error (NRMSE) between bright field images. Two optical configurations are suggested for recording bright field and fluorescence images simultaneously or alternately. The method was demonstrated on simulated data, and better than 0.3 nm drift correction precision was achieved. It was also applied on dSTORM imaging of F-actins of 3T3 cell, and the quality of reconstructed super-resolution image was improved observably. This method does not require special hardware, extra labelling or markers, and no precision decline due to photobleaching. It can be applied as an add-on for SMLM setups and achieves sub-nanometer precision drift correction for post-measurement or real time drift compensation. (C) 2014 Optical Society of America
Keywords:
PHOTOACTIVATION LOCALIZATION MICROSCOPY
OPTICAL MICROSCOPE
FLUORESCENCE
RECONSTRUCTION
REGISTRATION
NM
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