Open AccessThree-Dimensional Super-Resolved Imaging of Paraffin-Embedded Kidney Samples
Author(s) -
David UnnersjöJess,
Amer Ramdedovic,
Martin Höhne,
Linus Butt,
Felix C. Koehler,
RomanUlrich Müller,
Peter F. Hoyer,
Hans Blom,
Bernhard Schermer,
Thomas Benzing
Publication year - 2022
Publication title -
kidney360
Language(s) - English
Resource type - Journals
ISSN - 2641-7650
DOI - 10.34067/kid.0005882021
Subject(s) - podocyte , confocal , kidney disease , pathology , kidney , confocal microscopy , microscopy , filtration (mathematics) , gold standard (test) , biomedical engineering , biology , medicine , microbiology and biotechnology , radiology , optics , mathematics , proteinuria , statistics , physics
Background Diseases of the glomeruli, the renal filtration units, are a leading cause of progressive kidney disease. Assessment of the ultrastructure of podocytes at the glomerular filtration barrier is essential for diagnosing diverse disease entities, providing insight into the disease pathogenesis, and monitoring treatment responses. Methods Here we apply previously published sample preparation methods together with stimulated emission depletion and confocal microscopy for resolving nanoscale podocyte substructure. The protocols are modified and optimized in order to be applied to formalin-fixed paraffin-embedded (FFPE) samples. Results We successfully modified our protocols to allow for deep three-dimensional stimulated emission depletion and confocal imaging of FFPE kidney tissue with similar staining and image quality compared with our previous approaches. We further show that quantitative analysis can be applied to extract morphometrics from healthy and diseased samples from both mice and humans. Conclusions The results from this study could increase the feasibility of implementing optical kidney imaging protocols in clinical routines because FFPE is the gold-standard method for storage of patient samples.