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Image post-processing for SILMAS: structured illumination light sheet microscopy with axial sweeping.
David Frantz1, Courtney J Wright2, Allison J Schaser3
1Division of Combustion Physics, Department of Physics, Lund University, Lund, Sweden.
Biomedical Optics Express
|September 30, 2024
Summary
We developed a post-processing method to reduce artifacts in Scanning Light Sheet Microscopy (SILMAS) data. This technique enhances image quality for volumetric imaging of cleared tissues, improving quantitative analysis.
Area of Science:
- Biomedical Imaging
- Microscopy Techniques
- Computational Imaging
Background:
- Volumetric microscopy techniques like SILMAS offer improved contrast and optical sectioning.
- Efficient scattered light removal in SILMAS can accentuate artifacts like inhomogeneity, shadowing, and signal attenuation.
Purpose of the Study:
- To propose and validate a post-processing scheme for artifact suppression in SILMAS data.
- To enhance quantitative data extraction from volumetric microscopy of biological samples.
Main Methods:
- Applied a post-processing scheme combining Beer-Lambert law for light attenuation compensation and computational strategies for shadowing stripe suppression.
- Incorporated an additional bandpass step into wavelet-Fourier filtering for artifact removal without introducing new distortions.
- Demonstrated the technique on alpha-synuclein transgenic mouse brain data.
Main Results:
- Successfully suppressed illumination inhomogeneity, shadowing stripes, and signal attenuation in SILMAS volumetric data.
- Achieved near-quantitative compensation for light attenuation.
- Filtered high-contrast light sheet microscopy data with minimal distortion and no new artifacts.
Conclusions:
- The proposed post-processing scheme significantly improves the uniformity and quality of SILMAS volumetric data.
- The technique is suitable for imaging cleared tissue samples up to centimeter scale with micron-level isotropic resolution.
- This artifact suppression method enables more reliable quantitative analysis in volumetric microscopy.
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