Depth-variant deconvolution applied to widefield microscopy for rapid large-volume tissue imaging.

Area of Science:

• Biomedical Imaging
• Microscopy Techniques
• Tissue Engineering

Background:

• 3D tissue imaging is vital but hindered by long protocols and limited equipment access.
• Widefield microscopy offers speed and accessibility but lacks optical sectioning for 3D analysis.
• Existing methods often exclude widefield microscopy from advanced 3D imaging workflows.

Purpose of the Study:

• To develop an accessible 3D imaging method using widefield microscopy and deconvolution for thick tissues.
• To achieve subnuclear axial resolution in cleared tissues up to 500 µm deep.
• To demonstrate the method's utility in disease models and clinical evaluations.

Main Methods:

• Combined tissue clearing with an optimized commercial depth-variant deconvolution approach.
• Implemented prefiltering with z-brick splitting for enhanced axial resolution.
• Applied the method to large-volume, multi-tile widefield imaging of cleared tissues.

Main Results:

• Achieved subnuclear axial resolution in tissues up to 500 µm deep.
• Successfully visualized amyloid deposits in brain vasculature and atrophic tubules in kidney biopsies.
• Demonstrated resolution comparable to confocal microscopy in thick tissue sections.

Conclusions:

• Coupling widefield microscopy of cleared tissue with deconvolution provides an accessible method for 3D insight.
• This approach enhances research capabilities and facilitates clinical evaluations of thick tissue samples.
• The optimized method overcomes limitations of traditional 3D imaging, offering a viable alternative.