The method of morphometry study of kidney scarring has been described in previous studies.22 (link),23 (link) Briefly, Jones’ silver staining was performed on 5 μm thick sections of kidney biopsy specimen. Semiquantitative computerized image analysis was performed with the Leica Twin Pro image analysis system (Leica Microsystems), which was connected to a Leica DC500 digital camera on a Leica DMRXA2 microscope with a ×40 objective (final calibration: 0.258 mm/pixel). Image analysis was performed with MetaMorph 4.0 image-analyzing software (Universal Imaging Corporation). Ten glomeruli and 10 randomly selected areas were assessed in each patient’s sample and the average percentage of scarred glomerular and tubulointerstitial areas, as represented by the area with positive silver staining, were computed.
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4 protocols using twin pro
1
Morphometric Analysis of Kidney Scarring
2
Renal Scarring Morphometry Protocol
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The method of morphometry study of renal scarring has been described in previous studies [17, 18] (link). Briefly, Jones’ silver staining was performed on 5 μm thick sections of renal biopsy specimen. Semi-quantitative computerized image analysis was performed with the Leica Twin Pro image analysis system (Leica Microsystems, Wetzlar, Germany), which was connected to a Leica DC500 digital camera on a Leica DMRXA2 microscope with a ×40 objective (final calibration: 0.258 mm/pixel). Image analysis was performed by MetaMorph 4.0 image-analyzing software (Universal Imaging Corporation™, Downingtown, PA, USA). Ten glomeruli and 10 randomly selected areas were assessed in each patient, and the average percentage of scarred glomerular and tubulointerstitial areas, as represented by the area with positive silver staining, were computed.
3
Quantifying Nephrosclerosis in Renal Biopsies
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Jones’ silver staining was performed on 5 µm thick sections of the renal biopsy specimens of each patient. As previously described by others [34 (link)], we used a computerized image analysis method to semi-quantify nephrosclerosis. Briefly, a Leica Twin Pro image analysis system (Leica Microsystems, Wetzlar, Germany) was connected to a Leica DC500 digital camera on a Leica DMRXA2 microscope working with a 40× objective (final calibration: 0.258 mm/pixel), which was connected to a microcomputer for storage of the morphometric measurements so that image analysis could be performed using image analysis software (MetaMorph 4.0; Universal Imaging Corporation TM, Downingtown, PA, USA). A total of 10 glomeruli and 10 randomly selected areas were assessed in each patient and the average percentage of scarred glomerular and tubulointerstitial areas, as represented by the percentage of the area with positive staining, was computed for each patient.
4
Renal Scarring Quantification Protocol
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Renal scarring was studied using morphometry in previous studies [14 (link), 15 (link)]. Specifically, renal biopsy specimens that were 5 μm thick underwent Jones’ silver staining. Semi-quantitative computerized image analysis was then performed using the Leica Twin Pro image analysis system from Leica Microsystems in Wetzlar, Germany. The analysis utilized MetaMorph 4.0 image-analyzing software from Universal Imaging Corporation in Downingtown, PA, USA. Each patient’s sample included ten glomeruli and ten randomly selected areas, which were evaluated to determine the average percentage of scarred glomerular and tubulointerstitial areas.
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