Hjerpe A, Boon M E, Kok L P
Department of Pathology II, Karolinska Institute, Huddinge University Hospital, Sweden.
Histochem J. 1988 Jun-Jul;20(6-7):388-96. doi: 10.1007/BF01002733.
The effects of microwave stimulation on different aspects of immunohistochemical reactions were studied using Elisa as a model system. This technique allows monitoring of (i) the rates and recoveries of different antigen-antibody reactions, (ii) the formation of avidin/biotin complexes, (iii) the prevention of non-specific activity and (iv) the washing procedures. The binding reactions seemed to follow second-order kinetics, and reaction rates were considerably enhanced by microwave stimulation. The steps were aimed at preventing non-specific reactivity and the washing procedures could all be shortened to seconds. Such rate increases are far too large to be explained solely by the modest increase in temperature. Furthermore, microwave irradiation caused a major reduction in the yield of antigen-antibody complexes. This inhibitory effect can be compensated for by increasing the concentration of antibody. No significant effects of the microwaves on the antigen-antibody complexes were found once the complexes had been formed. The obtained results were then applied to the demonstration of carcinoembryonic antigen in histological material. By employing this method a complete immunohistochemical staining of a hydrated tissue section could be performed within 15 min, excluding the final development of colours.
以酶联免疫吸附测定(ELISA)作为模型系统,研究了微波刺激对免疫组织化学反应不同方面的影响。该技术能够监测:(i)不同抗原 - 抗体反应的速率和恢复情况;(ii)抗生物素蛋白/生物素复合物的形成;(iii)非特异性活性的抑制;以及(iv)洗涤过程。结合反应似乎遵循二级动力学,微波刺激可显著提高反应速率。这些步骤旨在防止非特异性反应,并且所有洗涤过程都可缩短至数秒。如此大幅度的速率增加,仅靠适度的温度升高是无法解释的。此外,微波辐射会导致抗原 - 抗体复合物的产量大幅降低。这种抑制作用可通过增加抗体浓度来补偿。一旦复合物形成,未发现微波对抗原 - 抗体复合物有显著影响。然后将所得结果应用于组织学材料中癌胚抗原的检测。采用这种方法,在不包括最终显色的情况下,15分钟内即可完成水合组织切片的完整免疫组织化学染色。