Three-dimensional microscopic analysis of clinical prostate specimens.

Area of Science:

• Biomedical imaging
• Pathology
• Cancer research

Background:

• Standard 2D prostate tissue sections limit understanding of 3D architecture.
• Three-dimensional (3D) architectural features of benign prostate tissue and prostate cancer (PCa) remain largely unknown.
• Investigating the 3D context is crucial for comprehensive analysis.

Purpose of the Study:

• To demonstrate the successful application of tissue-clearing protocols and confocal microscopy for 3D prostate tissue investigation.
• To overcome the limitations of 2D imaging in understanding prostate tissue architecture.
• To enable detailed visualization of benign and cancerous prostate structures in 3D.

Main Methods:

• Optical clearing of fresh and formalin-fixed paraffin-embedded (FFPE) prostate specimens.
• Confocal microscopy for deep tissue visualization (up to 800 μm).
• Fluorescent labeling with nuclear dye and cytokeratin (CK) 5, CK8-18 antibodies.

Main Results:

• Enabled visualization of prostate tissue structures up to 800 μm depth, a significant improvement over 70 μm in uncleared tissue.
• Provided comprehensive 3D imaging of benign prostate zones and individual PCa growth patterns.
• Preserved tissue integrity for subsequent 2D (immuno)histochemical staining and DNA analysis.

Conclusions:

• 3D imaging of clinical prostate specimens (fresh and FFPE) is feasible and highly valuable.
• This technique offers extensive opportunities for studying clinical and biological aspects in their true 3D context.
• Opens new avenues for research and diagnostics in prostate cancer and benign prostate diseases.