Dfc 480 digital camera

Paraffin embedded sections (5µm) were used for Masson’s trichrome staining and examined by a light microscope (Olympus photomicroscope linked to a DFC 480 digital camera). Semiquantitative scoring of the tubulointerstitial index was performed by random selection of 10 non-overlapping images of the kidney cortex using the following scoring system: score 0 = normal; score1 = mild interstitial fibrosis (≤ 10%); score2 = moderate interstitial fibrosis (10–25%); score3 = severe interstitial fibrosis (25–75%); score4 = extremely severe interstitial fibrosis (≥ 75%). The average scores from two blinded examiners were used for final analysis. The characteristics features for tubulointerstitial fibrosis included tubular atrophy or dilatation, presence of mononuclear inflammatory cells, widening of interstitial spaces with deposition of extracellular matrix, and interstitial cell proliferation.

Paraffin-embedded kidney sections were stained with Periodic Acid Schiff (PAS). The whole kidney cortex was examined under the magnification of 400x using a light microscope (Olympus photomicroscope linked to a DFC 480 digital camera). Two independent assessors, an anatomical pathologist and nephrologist, reviewed histological sections in a blinded manner and scored tubulointerstitial fibrosis, tubular injury and glomerulosclerosis. There was high inter- and intra-observer agreement for histological analysis between observers; and an average of 20 individual scores was calculated to generate the overall glomerulosclerosis score.
The characteristic features of tubulointerstitial fibrosis include tubular atrophy or dilatation, presence of mononuclear inflammatory cells, widening of interstitial spaces with deposition of extracellular matrix, interstitial cell proliferation and wrinkling or thickened tubular basement membrane (perivascular and periglomerular areas were discounted). Tubular interstitial fibrosis was scored as described in S1 Fig. Further analyses of tubular injury were assessed by: (A) tubular dilatation, (B) tubular vacuolation, C) glycogenated nuclei, and (D) tubular casts and scoring was performed as described in S2 Fig. Glomerulosclerosis was scored as previously described [12 (link), 16 (link)] and detailed in S3 Fig.