Evaluation by scanning acoustic microscopy (SAM) on glomerular lesion of IgA nephropathy

Abstract

IgA nephropathy (IgAN) is known to commonly cause of end‐stage renal failure in Japan. The glomerular lesions of IgAN have histological variations. The determination of prognosis and therapeutic strategy should be carefully done by experts because morphological information from renal biopsies using ordinary optical microscopy is usually qualitative and subjective. Moreover, the histological items for the evaluation of glomerular lesions seems to be unsatisfactory for expression of the disease condition of IgAN. The beneficial properties of scanning acoustic microscopy (SAM) include not only observation of microstructure but also quantitative measurement of acoustic propagation speed (APS), indicating the tissue elasticity. In the present study we compared the APS of glomeruli with the pathological scores that were determined by ordinary light microscopy. We used stocked human renal biopsy specimens diagnosed as IgAN (n = 12) and normal/minimal changes (n = 5). All samples were taken by renal biopsy in Kagawa Medical University Hospital during 1997–2000 under informed consent of the patients. The obtained renal tissue were immersed in 10% formalin and embedded in paraffin. A fixed specimen was consecutively cut into 4 μm slices. One of the deparaffinized 4 μm‐specimens was directly utilized for SAM without any staining, and the others were stained with haematoxylin‐eosin and Masson Trichrome for counting cell number and evaluation of collagen accumulation. For the measurement of glomerular APS, the sample line was set on the equator of the glomerulus and then scanning of the X–Z axis was carried out to obtain the interference fringes that were analysed with a computer imaging software in order to calculate the APS. In light microscopic study, pathological scores were evaluated semiquantitatively by two independent investigators who were unaware of the sample number. Glomerular lesions were scored into five grades and glomerular cell number was also counted in individual glomerulus. The computer‐assisted imaging analyser Win ROOF (Mitani, Fukui, Japan) was also used for the determination of glomerular collagen content in specimens stained by Masson Trichrome. A two‐dimensional image (C‐mode scanning) of SAM enabled imaging of glomerulus in renal biopsy specimen compatible with findings of ordinary light microscopy without staining dye. The glomerular APS in IgAN was significantly higher than in normal/minimal changes. This alteration of glomerular APS in IgAN was positively correlated to both semiquantitative pathological scores and glomerular collagen content determined by light microscopy. However, the cell number of glomelurus did not change between IgAN and normal/minimal change. As a result, we conclude that the glomerular lesion, especially matrix expansion in IgAN, was comparable with the absolute value among specimens. Therefore, it is suggested that SAM method is a novel and useful technique for quantitative evaluation of glomerular lesion in IgAN.