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首页> 美国卫生研究院文献>Clinical Molecular Pathology >Demonstration of immunoglobulin in cryostat and paraffin sections of human tonsil by immunofluorescence and immunoperoxidase techniques. Effects of processing on immunohistochemical performance of tissues and on the use of proteolytic enzymes to unmask antigens in sections.
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Demonstration of immunoglobulin in cryostat and paraffin sections of human tonsil by immunofluorescence and immunoperoxidase techniques. Effects of processing on immunohistochemical performance of tissues and on the use of proteolytic enzymes to unmask antigens in sections.

机译：通过免疫荧光和免疫过氧化物酶技术在人扁桃体的低温恒温器和石蜡切片中演示免疫球蛋白。加工对组织免疫组织化学性能的影响以及对蛋白水解酶用于揭露切片中抗原的影响。

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摘要

A fluorescein isothiocyanate (FITC) technique and one based on peroxidase-antiperoxidase (PAP) were used to study the distribution of immunoglobulin (Ig) in cryostat and paraffin sections of human tonsil. Trypsin and other proteolytic enzymes were used to 'unmask' the antigen in paraffin sections. The effects of processing, and particularly of fixation, on the immunohistochemical response of tissues were studied. The FITC and PAP methods detected Ig in paraffin and cryostat sections equally well. The distribution of the antigen was the same with both methods but the PAP method was the more informative. Formaldehyde-sucrose solution proved more suitable for fixing tissues for immunohistochemistry than glutaraldehyde. Trypsin revealed antigen in parraffin sections more efficiently than pepsin, papain, or pronase. Surface Ig (s-Ig) could be demonstrated in trypsinised paraffin sections but less effectively than in cryostat sections. Trypsinised paraffin sections were, however, more suitable for intracellular Ig (c-Ig) than cryostat sections although the performance of cryostat sections could be improved by prior fixation with a coagulative fixative.
机译：异硫氰酸荧光素（FITC）技术和基于过氧化物酶-抗过氧化物酶（PAP）的技术被用于研究免疫球蛋白（Ig）在人扁桃体的低温恒温器和石蜡切片中的分布。胰蛋白酶和其他蛋白水解酶用于“揭露”石蜡切片中的抗原。研究了加工过程，特别是固定过程对组织免疫组织化学反应的影响。 FITC和PAP方法在石蜡切片和低温恒温器切片中检测到的Ig效果相同。两种方法的抗原分布相同，但PAP方法的信息量更大。事实证明，甲醛-蔗糖溶液比戊二醛更适合固定组织用于免疫组织化学。胰蛋白酶比胃蛋白酶，木瓜蛋白酶或链霉蛋白酶更有效地揭示石蜡切片中的抗原。表面Ig（s-Ig）可以在胰蛋白酶处理的石蜡切片中显示，但效果不如低温恒温器切片。胰蛋白酶石蜡切片比冷冻切片机更适合细胞内Ig（c-Ig），尽管冷冻切片机的性能可以通过事先用凝固性固定剂固定来改善。

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