ArticleNature protocols2021
3D active stabilization for single-molecule imaging.
Article in Nature protocols, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 25 citations in OpenAlex.
- High-Science advances · 2026Article
- Nanomaterials and exercise interventions: A synergistic approach for atherosclerosis therapy (Review).International journal of molecular medicine · 2026Review
- Image-based 3D active sample stabilization on the nanometer scale for optical microscopy.Biophysical reports · 2025Article
- Open-source 3D active sample stabilization for fluorescence microscopy.Biophysical reports · 2025Article
- A Versatile Drift-Free Super-Resolution Imaging Method via Oblique Bright-Field Correlation.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- A practical guide to light-sheet microscopy for nanoscale imaging: Looking beyond the cell.Journal of applied physics · 2024Article
- The K2: Open-source simultaneous triple-color TIRF microscope for live-cell and single-molecule imaging.HardwareX · 2023Article
- Real-Time Feedback-Driven Single-Particle Tracking: A Survey and Perspective.Small (Weinheim an der Bergstrasse, Germany) · 2022Review
- Direct-laser writing for subnanometer focusing and single-molecule imaging.Nature communications · 2022Article
- A mean shift algorithm for drift correction in localization microscopy.Biophysical reports · 2021Article
- Building a Total Internal Reflection Microscope (TIRF) with Active Stabilization (Feedback SMLM).Bio-protocol · 2021Article
- A Tribute to Professor Katharina Gaus.Frontiers in bioinformatics · 2021Article
- Probing Biosensing Interfaces With Single Molecule Localization Microscopy (SMLM).Frontiers in chemistry · 2021Review
Corrections and comments
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Authors and funding
5 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A key part of any super-resolution technique involves accurately correcting for mechanical motion of the sample and setup during acquisition. If left uncorrected, drift degrades the resolution of the final reconstructed image and can introduce unwanted artifacts. Here, we describe how to implement active stabilization, thereby reducing drift to ~1 nm across all three dimensions. In this protocol, we show how to implement our method on custom and standard microscopy hardware. We detail the construction of a separate illumination and detection path, dedicated exclusively to acquiring the diffraction pattern of fiducials deposited on the imaging slide. We also show how to focus lock and adjust the focus in arbitrary nanometer step size increments. Our real-time focus locking is based on kHz calculations performed using the graphics processing unit. The fast calculations allow for rapid repositioning of the sample, which reduces drift below the photon-limited localization precision. Our approach allows for a single-molecule and/or super-resolution image acquisition free from movement artifacts and eliminates the need for complex algorithms or hardware installations. The method is also useful for long acquisitions which span over hours or days, such as multicolor super resolution. Installation does not require specialist knowledge and can be implemented in standard biological laboratories. The full protocol can be implemented within ~2 weeks.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.