High-Fidelity Imaging in Brain-Wide Structural Studies Using Light-Sheet Microscopy.

Area of Science:

• Neuroscience
• Optical Imaging
• Biophysics

Background:

• Light-sheet microscopy (LSM) is crucial for high-resolution, high-speed imaging of whole brains, especially with tissue clearing.
• Opaque objects in cleared samples cause stripe artifacts in LSM, degrading image homogeneity and obscuring neural and vascular structures.
• Gaussian illumination, commonly used in LSM, is susceptible to these artifacts, limiting data quality.

Purpose of the Study:

• To investigate the efficacy of Bessel beams in reducing stripe artifacts in light-sheet microscopy of intact, optically cleared mouse brains.
• To assess the impact of Bessel beams on image fidelity and information content compared to Gaussian illumination.
• To evaluate the utility of Bessel beams for reconstructing brain-wide neuronal and vascular network morphology.

Main Methods:

• Light-sheet microscopy was employed on intact, optically cleared mouse brains.
• Bessel beams were utilized as an illumination source and compared against Gaussian illumination.
• Image quality, artifact reduction, and accessibility of information content were analyzed in whole-brain datasets.

Main Results:

• Bessel beams significantly reduced stripe artifacts, leading to improved image homogeneity.
• Approximately one-third of the imaged brain volume was affected by severe artifacts with conventional illumination.
• Bessel beams recovered significant information content lost due to artifacts with Gaussian illumination, enhancing visualization of neuronal and vascular networks.

Conclusions:

• Bessel beams provide high-fidelity structural data with enhanced image homogeneity in light-sheet microscopy of cleared mouse brains.
• This method improves the visualization of brain-wide neuronal and vascular morphology.
• The use of Bessel beams may simplify automated analysis of neural and vascular features by reducing artifacts.