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Related Experiment Video

Updated: Jan 1, 2026

Sample Drift Correction Following 4D Confocal Time-lapse Imaging

Sample Drift Correction Following 4D Confocal Time-lapse Imaging

Published on: April 12, 2014

Sample drift estimation method based on speckle patterns formed by backscattered laser light.

Shih-Ya Chen¹, Rainer Heintzmann^2,3, Christoph Cremer^1,4,5

¹Institute of Molecular Biology, Mainz, Germany.

Biomedical Optics Express

|December 20, 2019

Summary

This summary is machine-generated.

We developed a new method to correct sample drift during long single-molecule localization microscopy (SMLM) experiments. This technique uses scattered laser light to achieve high-precision 3D drift correction without extra sample preparation.

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Last Updated: Jan 1, 2026

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Area of Science:

Biophysics
Microscopy techniques
Optical imaging

Background:

Single molecule localization microscopy (SMLM) achieves nanoscale resolution but is limited by sample drift during long acquisition times.
Accurate drift estimation is crucial for high-resolution SMLM imaging over extended periods.

Purpose of the Study:

To present a novel method for precise 3D drift correction in SMLM.
To enable accurate long-term SMLM measurements by mitigating sample movement artifacts.

Main Methods:

Utilized speckle patterns from backscattered laser light within the SMLM setup.
Recorded a 3D reference stack of unique speckle patterns before measurements.
Correlated real-time scattered light images with the reference stack to determine x, y, and z drift.

Main Results:

Achieved drift correction precision comparable to fiducial marker methods, reaching several nanometers.
Demonstrated effective 3D drift estimation and correction for SMLM.
Validated the method's performance in correcting sample drift.

Conclusions:

The speckle pattern evaluation method provides high-precision 3D drift correction for SMLM.
This technique eliminates the need for additional sample preparation for drift correction.
Enables more robust and reliable long-term SMLM imaging.