A New Quantitative Metric for Precise Classification of Diabetic Podocyte Injury Using Scanning Electron Microscopy

Faith Rooney; Chantal Allamargot; Jillian Williquett; Hua Sun

doi:10.1093/mam/ozaf122

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A New Quantitative Metric for Precise Classification of Diabetic Podocyte Injury Using Scanning Electron Microscopy

Journal article

Open access

Peer reviewed

A New Quantitative Metric for Precise Classification of Diabetic Podocyte Injury Using Scanning Electron Microscopy

Faith Rooney, Chantal Allamargot, Jillian Williquett and Hua Sun

Microscopy and microanalysis, Vol.31(6), ozaf122

11/12/2025

DOI: 10.1093/mam/ozaf122

PMCID: PMC12693405

PMID: 41370191

Appears in UI Libraries Support Open Access

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Abstract

Podocytes are highly interdigitated epithelial cells in the glomerulus that maintain the kidney's filtration barrier, and their injury underlies the progression of diabetic kidney disease. Early podocyte damage is challenging to detect using light or transmission electron microscopy; new techniques like super-resolution microscopy remain limited in capturing the three-dimensional topography of podocytes. Scanning electron microscopy (SEM) offers superior spatial resolution and surface detail; however, standardized quantitative methods to analyze podocyte ultrastructure are lacking. In this study, we developed and compared three analytical approaches to quantify podocyte injury from SEM images in a streptozotocin-induced diabetic mouse model. Using ImageJ software, we measured the slit diaphragm (SD) fraction via (1) thresholding, (2) ridge detection, and (3) foot process plot profiling, comparing diabetic and nondiabetic podocytes. The ridge detection method showed the best diagnostic accuracy (88% sensitivity and 93% specificity), successfully distinguishing diabetic from healthy podocytes. Furthermore, SD fraction measurements correlated negatively with biomarkers of podocyte dysfunction and diabetic stress. This work establishes the first reliable, quantitative pipeline for detecting subtle early podocyte injury in diabetic kidney disease using SEM, providing a valuable tool for future mechanistic and therapeutic studies.

Animals

Diabetes Mellitus, Experimental - pathology

Diabetic Nephropathies - pathology

Disease Models, Animal

Image Processing, Computer-Assisted - methods

Male

Mice

Mice, Inbred C57BL

Microscopy, Electron, Scanning - methods

Podocytes - pathology

Podocytes - ultrastructure

Sensitivity and Specificity

UIOWA OA Agreement

Details

Title: Subtitle: A New Quantitative Metric for Precise Classification of Diabetic Podocyte Injury Using Scanning Electron Microscopy
Creators: Faith Rooney - University of Iowa
Chantal Allamargot - University of Iowa
Jillian Williquett - University of Iowa, Stead Family Department of Pediatrics
Hua Sun - University of Iowa
Resource Type: Journal article
Publication Details: Microscopy and microanalysis, Vol.31(6), ozaf122
DOI: 10.1093/mam/ozaf122
PMID: 41370191
PMCID: PMC12693405
NLM abbreviation: Microsc Microanal
ISSN: 1431-9276
eISSN: 1435-8115
Publisher: Oxford University Press
Grant note: Roy J. and Lucille A. Carver College of Medicine CMN86023539 / University of Iowa R01DK136563 / NIDDK NIH HHS
Language: English
Date published: 11/12/2025
Academic Unit: Nephrology, Dialysis and Transplantation; Stead Family Department of Pediatrics; Core Research Facilities
Record Identifier: 9985091811802771

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