Costa M, Buffa R, Furness J B, Solcia E
Histochemistry. 1980 Feb;65(2):157-65. doi: 10.1007/BF00493164.
A method is described for the immunohistochemical localization of peptides in whole-mount preparations. Tissue is fixed as laminae with a picric acid/formaldehyde mixture and then dehydrated, cleared and rehydrated before exposure to antibodies. This procedure ensures adequate penetration of the antibody molecules without the need to freeze and thaw the tissue or to use detergents, preserves antigenicity and lowers non-specific background staining. The laminae are incubated with the primary antisera for 16 h at room temperature and, after washing, with a second, fluorescent tagged, antiserum. This can be followed by a peroxidase-anti-peroxidase localization of the second antiserum, which acts as a bridge. The method gives a precise and reproducible localization of immunoreactive peptides, with good penetration and low background even in thick preparation. Large areas can be scanned and neuroeffector relationships studied more easily than in sections.
本文描述了一种在整装标本中对肽进行免疫组织化学定位的方法。组织用苦味酸/甲醛混合物固定成薄片，然后在暴露于抗体之前进行脱水、透明和再水化。该程序可确保抗体分子充分渗透，而无需对组织进行冻融或使用去污剂，保留了抗原性并降低了非特异性背景染色。将薄片在室温下与一抗血清孵育16小时，洗涤后，再与第二种荧光标记的抗血清孵育。随后可以对作为桥梁的第二种抗血清进行过氧化物酶-抗过氧化物酶定位。该方法能对免疫反应性肽进行精确且可重复的定位，即使在厚标本中也具有良好的穿透力和低背景。与切片相比，可以更容易地扫描大面积区域并研究神经效应关系。
