Low-vacuum scanning electron microscopy imaging of kidney vascular endothelium using metal-sensitized immunochemistry

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  • BABA Masaki
    Department of Pathology, Tsukuba University Hospital
  • KAWANISHI Kunio
    Department of Pathology, Tsukuba University Hospital Department of Experimental Pathology, Faculty of Medicine, University of Tsukuba
  • NAKAGAWA Tomoki
    Department of Pathology, Tsukuba University Hospital
  • MURATA Yoshihiko
    Department of Pathology, Tsukuba University Hospital
  • FURUYA Shuichiro
    Department of Pathology, Tsukuba University Hospital
  • MATSUBARA Daisuke
    Department of Pathology, Tsukuba University Hospital Department of Diagnostic Pathology, Faculty of Medicine, University of Tsukuba
  • KATO Mitsuyasu
    Department of Experimental Pathology, Faculty of Medicine, University of Tsukuba

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  • 免疫組織化学後の3,3'-ジアミノベンジジン四塩酸塩水和物の金属増感法を用いた腎血管内皮の低真空走査型電子顕微鏡解析
  • メンエキ ソシキ カガク ゴ ノ 3,3'-ジアミノベンジジン ヨン エンサンエン スイワブツ ノ キンゾク ゾウカンホウ オ モチイタ ジンケッカン ナイヒ ノ テイシンクウ ソウサガタ デンシ ケンビキョウ カイセキ

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Abstract

<p>The pathological diagnosis of renal needle biopsy specimens requires the use of various analytical methods, including the use of various special stains, immunofluorescence, immunohistochemistry, and transmission electron microscopy (TEM). Inevitably, there are many cases wherein it is difficult to make a definitive diagnosis because of insufficient materials, especially for TEM. To compensate for this, low-vacuum scanning electron microscopy (LVSEM) imaging using formalin-fixed paraffin-embedded (FFPE) specimens is being tested as a possible diagnostic tool for kidney biopsy. Here, we report a new LVSEM imaging technique using metal-sensitized immunohistochemistry. We performed immunohistochemistry on FFPE samples of kidney tissue to detect CD31 and CD34, which are used as vascular endothelial markers. We then sensitized 3,3'-diaminobenzidine (DAB) with AuCl. We also performed platinum blue and thiosemicarbazide-periodic acid-methenamine silver (TSC-PAM) staining for the control of LVSEM imaging as previously reported. LVSEM imaging of CD31 and CD34 clearly visualized glomerular endothelial cells (GECs), the peritubular capillary network, and small arteries, which are difficult to identify in the case of platinum blue or TSC-PAM staining, suggesting that LVSEM imaging can be useful in the assessment of inflammatory disease and tumor invasion. In conclusion, we established a new LVSEM imaging technique for kidney endothelium, and the AuCl sensitization of DAB can be applied to any type of immunohistochemistry to observe microstructures as well as to confirm the deposition of immune complexes in renal biopsy specimens.</p>

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