Three-Dimensional Super-Resolved Imaging of Paraffin-Embedded Kidney Samples

Area of Science:

• Nephrology
• Microscopy
• Biomedical Imaging

Background:

• Glomerular diseases are a primary cause of progressive kidney disease.
• Assessing podocyte ultrastructure is crucial for diagnosing kidney diseases, understanding pathogenesis, and monitoring treatment.
• Podocytes are key components of the glomerular filtration barrier.

Purpose of the Study:

• To adapt and optimize nanoscale imaging techniques for podocyte substructure analysis.
• To enable the application of stimulated emission depletion (STED) and confocal microscopy to formalin-fixed paraffin-embedded (FFPE) kidney samples.
• To facilitate the quantitative analysis of podocyte morphometrics in both healthy and diseased kidney tissues.

Main Methods:

• Utilized stimulated emission depletion (STED) and confocal microscopy.
• Modified and optimized established sample preparation protocols.
• Applied methods to formalin-fixed paraffin-embedded (FFPE) kidney tissue samples.

Main Results:

• Successfully adapted protocols for deep three-dimensional imaging of FFPE kidney tissue.
• Achieved comparable staining and image quality to previous methods using fresh-frozen samples.
• Demonstrated the feasibility of quantitative morphometric analysis on FFPE samples from mice and humans.

Conclusions:

• Optimized optical imaging protocols can be applied to FFPE kidney samples.
• This advancement enhances the feasibility of integrating advanced optical kidney imaging into clinical practice.
• FFPE sample compatibility is key for routine clinical application due to established storage methods.