Litwin J A, Völkl A, Stachura J, Fahimi H D
Department of Histology, Copernicus Medical Academy, Kraków, Poland.
Histochem J. 1988 Mar;20(3):165-73. doi: 10.1007/BF01746680.
We describe the immunocytochemical localization of four peroxisomal enzymes by light microscopy in human liver and kidney processed routinely by formalin fixation and paraffin embedding. Monospecific antisera against catalase and three enzymes of peroxisomal lipid beta-oxidation (acyl-CoA oxidase, bifunctional protein (enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase) and 3-ketoacyl-CoA thiolase) were used in conjunction with either the indirect immunoperoxidase method or the protein A-gold technique followed by silver intensification. The sections of formalin-fixed paraffin-embedded tissue had to be deparaffinized and subjected to controlled proteolysis in order to obtain satisfactory immunostaining. Under the conditions employed, peroxisomes were distinctly visualized in liver parenchymal cells with no reaction in bile duct epithelial or sinusoidal lining cells. In the kidney, peroxisomes were confined to the proximal tubular epithelial cells with negative staining of glomeruli, distal tubules and collecting ducts. A positive immunocytochemical reaction was obtained even in paraffin blocks stored for several years. The method offers a simple approach for detection of peroxisomes and evaluation of their various enzyme proteins in material processed routinely in histopathology laboratories and should prove useful in the investigation of the role of peroxisomes in human pathology for both prospective and retrospective studies.
我们通过光学显微镜描述了四种过氧化物酶体酶在经福尔马林固定和石蜡包埋常规处理的人肝脏和肾脏中的免疫细胞化学定位。使用针对过氧化氢酶和过氧化物酶体脂质β-氧化的三种酶（酰基辅酶A氧化酶、双功能蛋白（烯酰辅酶A水合酶、3-羟酰基辅酶A脱氢酶）和3-酮酰基辅酶A硫解酶）的单特异性抗血清，结合间接免疫过氧化物酶法或蛋白A-金技术，随后进行银增强。福尔马林固定石蜡包埋组织的切片必须进行脱蜡并进行可控的蛋白水解，以获得满意的免疫染色。在所采用的条件下，过氧化物酶体在肝实质细胞中清晰可见，胆管上皮细胞或窦状衬里细胞无反应。在肾脏中，过氧化物酶体局限于近端肾小管上皮细胞，肾小球、远端小管和集合管呈阴性染色。即使在储存数年的石蜡块中也获得了阳性免疫细胞化学反应。该方法为在组织病理学实验室常规处理的材料中检测过氧化物酶体及其各种酶蛋白提供了一种简单的方法，并且在过氧化物酶体在人类病理学中的作用的前瞻性和回顾性研究中应该是有用的。
