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A Versatile Drift‐Free Super‐Resolution Imaging Method via Oblique Bright‐Field Correlation
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A Versatile Drift‐Free Super‐Resolution Imaging Method via Oblique Bright‐Field Correlation
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Journal Article
A Versatile Drift‐Free Super‐Resolution Imaging Method via Oblique Bright‐Field Correlation
2025
Overview
High‐resolution optical microscopy, particularly super‐resolution localization microscopy, requires precise real‐time drift correction to maintain constant focus at nanoscale precision during the prolonged data acquisition. Existing methods, such as fiducial marker tracking, reflection monitoring, and bright‐field image correlation, each provide certain advantages but are limited in their broad applicability. In this work, a versatile and robust drift correction technique is presented for single‐molecule localization‐based super‐resolution microscopy. It is based on the displacement analysis of bright‐field image features of the specimen with oblique illumination. By leveraging the monotonic relationship between the displacement of image features and axial positions, this method can precisely measure the drift of the imaging system in real‐time with sub‐nanometer precision in all three dimensions, over a broad axial range, and for various samples, including those with closely matched refractive indices. The performance of this method is validated against conventional marker‐assisted techniques and demonstrates its high precision in super‐resolution imaging across various biological samples. This method paves the way for fully automated drift‐free super‐resolution imaging systems. This study introduces a robust marker‐free drift correction method for single‐molecule localization‐based super‐resolution microscopy. Utilizing oblique illumination, it analyzes bright‐field image feature displacement to achieve sub‐nanometer precision in real‐time drift tracking across all dimensions. Validated against conventional techniques, this versatile approach supports automated drift‐free imaging, ensuring nanoscale precision for prolonged high‐resolution microscopy in diverse biological samples.
Publisher
John Wiley & Sons, Inc,John Wiley and Sons Inc
Subject
