High-resolution 3D fluorescent imaging of intact tissues

Area of Science:

• Histology and Molecular Imaging
• Biomedical Research Techniques

Background:

• Assessing molecular structures in large cells and human organs is challenging due to limited reagent and light penetration in opaque tissues.
• Existing methods struggle to balance high-resolution imaging with the spatial constraints of large tissue samples.

Purpose of the Study:

• To describe an optimized sample preparation method for sub-micron resolution 3D imaging of human and rodent tissues.
• To achieve greater imaging depth and resolution in intact organs compared to previous techniques.

Main Methods:

• Developed optimized sample preparation for cleared tissue sections (>100 microm thick).
• Utilized confocal microscopy to generate 3D images of adult human and rodent organs (heart, kidney, liver).
• Applied various chemical and antibody stains to labeled tissues.

Main Results:

• Achieved imaging depth greater than 100 micrometers.
• Obtained sub-micron resolution with a voxel size less than 0.012 microm³.
• Successfully imaged cleared adult human and rodent organs, including heart, kidney, and liver.

Conclusions:

• The described method enables high-resolution 3D imaging in thick tissue sections, previously limited to smaller model systems.
• This technique can be readily adopted by histology labs for routine use.
• The entire process from tissue preparation to 3D imaging takes approximately 3 days.