Related Experiment Video
Updated: Jun 25, 2026
Assessment of Kidney Function in Mouse Models of Glomerular Disease
Published on: June 30, 2018
Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal
Anand C Reghuvaran1, Qisheng Lin2, John M Basgen3
1Division of Nephrology, Department of Medicine, Yale University School of Medicine, New Haven, Connecticut, USA.
Abstract:
Morphometric estimates of mean or individual glomerular volume (MGV, IGV) have biological implications, over and above qualitative histologic data. However, morphometry is time-consuming and requires expertise limiting its utility in clinical cases. We evaluated MGV and IGV using plastic- and paraffin-embedded tissue from 10 control and 10 focal segmental glomerulosclerosis (FSGS) mice (aging and 5/6th nephrectomy models) using the gold standard Cavalieri (Cav) method versus the 2-profile and Weibel-Gomez (WG) methods and a novel 3-profile method. We compared accuracy, bias and precision, and quantified results obtained when sampling differing numbers of glomeruli. In both FSGS and controls, we identified an acceptable precision for MGV of 10-glomerular sampling versus 20-glomerular sampling using the Cav method, while 5-glomerular sampling was less precise. In plastic tissue, 2- or 3-profile MGVs showed greater concordance with MGV when using Cav, versus MGV with WG. IGV comparisons using the same glomeruli reported a consistent underestimation bias with both 2- or 3-profile methods versus the Cav method. FSGS glomeruli showed wider variations in bias estimation than controls. Our 3-profile method offered incremental benefit to the 2-profile method in both IGV and MGV estimation (improved correlation coefficient, Lin's concordance and reduced bias). In our control animals, we quantified a shrinkage artifact of 52% from tissue processed for paraffin-embedded versus plastic-embedded tissue. FSGS glomeruli showed overall reduced shrinkage albeit with variable artifact signifying periglomerular/glomerular fibrosis. A novel 3-profile method offers slightly improved concordance with reduced bias versus 2-profile. Our findings have implications for future studies using glomerular morphometry.
Insights
Glomerular volume estimation using a novel 3-profile method shows improved accuracy and reduced bias compared to existing techniques. This method offers a more precise way to assess kidney disease, even with fewer glomeruli samples.
Area of Science:
- Nephrology
- Quantitative Pathology
- Biomedical Engineering
Background:
- Morphometric estimation of glomerular volume (MGV, IGV) is crucial for understanding kidney disease but is often limited by time and expertise.
- Existing methods for glomerular morphometry, such as the Cavalieri (Cav) method, 2-profile, and Weibel-Gomez (WG) methods, have limitations in accuracy, bias, and precision.
- Focal segmental glomerulosclerosis (FSGS) is a significant kidney disease where glomerular morphometry can provide valuable insights.
Purpose of the Study:
- To evaluate and compare the accuracy, bias, and precision of different methods for estimating mean glomerular volume (MGV) and individual glomerular volume (IGV).
- To assess the impact of varying glomerular sampling numbers on the precision of MGV estimates.
- To introduce and validate a novel 3-profile method for glomerular morphometry and compare it with existing techniques.
Main Methods:
- Morphometric analysis of MGV and IGV was performed on plastic- and paraffin-embedded kidney tissue from control and FSGS mice using the Cavalieri (Cav) method, 2-profile, Weibel-Gomez (WG), and a novel 3-profile method.
- Accuracy, bias, and precision were compared across methods and with different numbers of glomeruli sampled (5, 10, 20).
- Tissue shrinkage artifacts between plastic- and paraffin-embedded samples were quantified.
Main Results:
- An acceptable precision for MGV estimation was achieved with 10-glomerular sampling using the Cav method, while 5-glomerular sampling was less precise.
- The 2- and 3-profile methods showed greater concordance with Cav-based MGV in plastic-embedded tissue compared to WG.
- A consistent underestimation bias was observed with 2- and 3-profile methods for IGV compared to the Cav method, with greater variability in FSGS glomeruli.
- The novel 3-profile method demonstrated incremental benefits over the 2-profile method, showing improved correlation, Lin's concordance, and reduced bias.
- A significant shrinkage artifact (52%) was quantified in paraffin-embedded versus plastic-embedded control tissues, with reduced but variable shrinkage in FSGS glomeruli.
Conclusions:
- The novel 3-profile method offers improved concordance and reduced bias for glomerular volume estimation compared to the 2-profile method.
- Glomerular morphometry, particularly with optimized sampling strategies, provides valuable biological data beyond qualitative histology.
- Understanding tissue processing artifacts, like shrinkage, is critical for accurate morphometric analysis in kidney disease studies.
