【摘要】 本文研究了不同固定液对于未标记抗体酶(PAP)技术显示石蜡包埋大鼠淋巴结切片中细胞内免疫球蛋白(IgG)和组织形态的作用。Bouin氏液对于IgG抗原性的保存为最好,染色界线清晰,组织形态保持完整。更多还原

【Abstract】 The effect of various fixatives on the immunoglobulin immunological activity of antigenic determinant sites of intracellular immunoglobulin has until lately only been studied to a limited extent. The present investigation has studied the effect of different fixatives on the demonstration of intraeellular immunoglobulin (IgG) and tissue morphology in paraffin embedded lymph nodes. Samples of lymph node were obtained from syngeneic WAB/Not male rats carried with hepatoma D23/ 47 and operated upon under ether inhalation anethesia.The right and left lumbar lymph nodes were fixed in fixatives including Bouin’s fluid, B-5, modified Millonig’s phosphatebuffered formalin, Zamboni’s fixa- tive and paraformaldehyde. The tissues were dehydrated in ethanol, cleared in zylene and embedded in paraffin.Staining was performed by PAP methods. Rabbit ant-irat IgG was used as first antibody, swine anti-rabbit IgG as second antibody, and soluble complex of horseradish peroxidase (HRP)-rabbit anti-horseradish peroxidase (PAP)as third antibody. The slides were incubated for peroxidase activity in3,3′-d iaminobenzidine tetrahydrochloride (DAB) medium. The greatest number of positively staining cells was seen in the cortex of rat lymph nodes. Bouin’s fluid produced good preservation of antigenicity of immunoglobulins,sharp demarcation of the staining reaction and adequate preservation of tissue morphology as well. But the other fixatives gave a less satifactory result with bad tissue morphology and/or intense background staining. The author suggests fixation in Bouin’s fluid so as to obtain a result demonstrating large amount of stained immunoglobulin containing celts and well preserved tissue morphology on paraffin embedded material.更多还原