Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruli
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‐ a...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-07-01
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| Series: | Physiological Reports |
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| Online Access: | https://doi.org/10.14814/phy2.15688 |
| _version_ | 1797397521240162304 |
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| author | Anand C. Reghuvaran Qisheng Lin John M. Basgen Khadija Banu Hongmei Shi Anushree Vashist John Pell Sudhir Perinchery John C. He Dennis Moledina F. Perry Wilson Madhav C. Menon |
| author_facet | Anand C. Reghuvaran Qisheng Lin John M. Basgen Khadija Banu Hongmei Shi Anushree Vashist John Pell Sudhir Perinchery John C. He Dennis Moledina F. Perry Wilson Madhav C. Menon |
| author_sort | Anand C. Reghuvaran |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T01:12:16Z |
| format | Article |
| id | doaj.art-02e05503c5ac4263a18f0e17dfc71189 |
| institution | Directory Open Access Journal |
| issn | 2051-817X |
| language | English |
| last_indexed | 2024-03-09T01:12:16Z |
| publishDate | 2023-07-01 |
| publisher | Wiley |
| record_format | Article |
| series | Physiological Reports |
| spelling | doaj.art-02e05503c5ac4263a18f0e17dfc711892023-12-11T03:20:34ZengWileyPhysiological Reports2051-817X2023-07-011113n/an/a10.14814/phy2.15688Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruliAnand C. Reghuvaran0Qisheng Lin1John M. Basgen2Khadija Banu3Hongmei Shi4Anushree Vashist5John Pell6Sudhir Perinchery7John C. He8Dennis Moledina9F. Perry Wilson10Madhav C. Menon11Division of Nephrology, Department of Medicine Yale University School of Medicine New Haven Connecticut USADepartment of Nephrology, Renji Hospital, School of Medicine Shanghai Jiao Tong University Shanghai People's Republic of ChinaMorphometry and Stereology Laboratory Charles R. Drew University of Medicine and Science Los Angeles California USADivision of Nephrology, Department of Medicine Yale University School of Medicine New Haven Connecticut USADivision of Nephrology, Department of Medicine Yale University School of Medicine New Haven Connecticut USADivision of Nephrology, Department of Medicine Yale University School of Medicine New Haven Connecticut USADivision of Nephrology, Department of Medicine Yale University School of Medicine New Haven Connecticut USADepartment of Pathology Yale University School of Medicine New Haven Connecticut USADivision of Nephrology, Department of Medicine Icahn School of Medicine at Mount Sinai New York New York USADivision of Nephrology, Department of Medicine Yale University School of Medicine New Haven Connecticut USADivision of Nephrology, Department of Medicine Yale University School of Medicine New Haven Connecticut USADivision of Nephrology, Department of Medicine Yale University School of Medicine New Haven Connecticut USAAbstract 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.https://doi.org/10.14814/phy2.156883‐Profile methodCavalieri methodglomerular morphometryWeibel–Gomez method |
| spellingShingle | Anand C. Reghuvaran Qisheng Lin John M. Basgen Khadija Banu Hongmei Shi Anushree Vashist John Pell Sudhir Perinchery John C. He Dennis Moledina F. Perry Wilson Madhav C. Menon Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruli Physiological Reports 3‐Profile method Cavalieri method glomerular morphometry Weibel–Gomez method |
| title | Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruli |
| title_full | Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruli |
| title_fullStr | Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruli |
| title_full_unstemmed | Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruli |
| title_short | Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruli |
| title_sort | comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruli |
| topic | 3‐Profile method Cavalieri method glomerular morphometry Weibel–Gomez method |
| url | https://doi.org/10.14814/phy2.15688 |
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