Stoward P J
Ciba Found Symp. 1979(73):11-31. doi: 10.1002/9780470720561.ch3.
Some practical criteria are suggested for establishing the precision, reproducibility, validity and specificity of quantitative histochemical techniques used for assaying the activities of enzymes in single cells and tissue sections. To be valid, a technique should ideally pass 12 tests. Principally these involve proving that the mean absorbance or fluorescence of the specific final reaction product (FRP) is related to section thickness, incubation time, substrate concentration and the concentration of enzyme in situ. However, the formation of appreciable amounts of non-specific FRP may interfere in the determination of the true enzyme activity. This and other difficulties are illustrated with data obtained from an investigation of Meijer's semipermeable membrane technique for assaying acid phosphatase in unfixed sections of muscle.
本文提出了一些实用标准，用于确立在单细胞和组织切片中测定酶活性的定量组织化学技术的精密度、重现性、有效性和特异性。一项技术若要有效，理想情况下应通过12项测试。这些测试主要包括证明特定最终反应产物（FRP）的平均吸光度或荧光与切片厚度、孵育时间、底物浓度以及原位酶浓度相关。然而，大量非特异性FRP的形成可能会干扰对真实酶活性的测定。通过对Meijer用于测定肌肉未固定切片中酸性磷酸酶的半透膜技术的研究获得的数据，说明了这一问题及其他困难。
