Related Experiment Video
Updated: Feb 6, 2026
Specific Labeling of Mitochondrial Nucleoids for Time-lapse Structured Illumination Microscopy
Published on: June 4, 2020
Automated distinction of shearing and distortion artefacts in structured illumination microscopy
Abstract:
Any motion during an image acquisition leads to an artefact in the final image. Structured illumination microscopy (SIM) combines several raw images into one high-resolution image and is thus particularly prone to these motion artefacts. Their unpredictable shape cannot easily be distinguished from real high-resolution content. We previously implemented a motion detection specifically for SIM, which had two shortcomings which are solved here. First, the brightness dependency of the motion signal is removed. Second, the empirical threshold of the calculated motion signal was not a threshold at a maximum allowed artefact. Here we investigate which artefacts are still acceptable and which linear movement creates them. Thus, the motion signal is linked with the maximal strength of the expected artefact. A signal-to-noise analysis including classification successfully distinguishes between artefact-free imaging, shearing and distortion artefacts in biological specimens. A shearing, as in wide-field microscopy, is the dominant reconstruction artefact, while distortions arise not until surprisingly fast movements.
Insights
This study refines motion detection for structured illumination microscopy (SIM), removing brightness dependency and linking motion signals to acceptable artifact levels for clearer high-resolution imaging.
Area of Science:
- Microscopy
- Image Processing
- Biophysics
Background:
- Motion during image acquisition causes artifacts in microscopy, particularly in structured illumination microscopy (SIM) due to image combination.
- Existing SIM motion detection methods have limitations, including brightness dependency and arbitrary artifact thresholds.
Purpose of the Study:
- To improve motion detection in SIM by eliminating brightness dependency.
- To establish a motion signal threshold directly related to acceptable artifact levels.
- To differentiate between artifact-free imaging, shearing, and distortion artifacts.
Main Methods:
- Developed a motion detection method for SIM without brightness dependency.
- Linked the motion signal strength to the maximum permissible artifact size.
- Employed signal-to-noise analysis and classification to categorize imaging artifacts.
Main Results:
- The enhanced method successfully distinguishes between artifact-free, shearing, and distortion artifacts.
- Shearing is identified as the primary reconstruction artifact in SIM, similar to wide-field microscopy.
- Distortion artifacts only emerge at significantly higher movement speeds.
Conclusions:
- The improved motion detection algorithm enhances the reliability of SIM by accurately identifying and classifying motion-induced artifacts.
- This advancement allows for better discrimination between true high-resolution content and image degradation in SIM.
- Understanding artifact types and their relation to movement speed is crucial for optimizing imaging protocols in super-resolution microscopy.
Related Concept Videos
01:27Shear Diagram
First, a free-body diagram of the beam is drawn, representing all the external forces and internal reactions acting on the beam. One can calculate the reaction forces at each support by employing the equilibrium equations of force and moment. The vertical component...
01:19Shearing Stress
The average shearing stress can be calculated by dividing the shear by the area of the cross-section.
01:20Shearing Strain
01:02¹H NMR: Interpreting Distorted and Overlapping Signals
As Δν decreases and the signals move closer, the doublets appear increasingly distorted. The intensities of the inner lines increase at the cost of those of the outer lines as the signals are...
02:34Structural Isomerism
Isomers are different chemical species that have the same chemical formula. Structural isomerism of coordination compounds can be divided into two subcategories, the linkage isomers and coordination-sphere isomers.
Linkage isomers occur when the coordination compound contains a ligand that can bind to the transition metal center through two different atoms. For example, the CN− ligand can bind through the carbon atom or through the nitrogen atom. Similarly, SCN− can...
01:14Normal and Shear Force