免疫组织化学染色定量方法研究（Ⅲ）

STUDY ON THE QUANTITATIVE METHOD OF IMMUNOHISTOCHEMISTRY(Ⅲ)

本文对免疫组织化学染色反应的定量方法进行了研究，导出阳性单位（ＰＵ）测算新公式。根据测试区域Ａ的灰度级ＧＡ和面积（ｍ＋ｎ）及阳性反应产物α相的灰度级Ｇα和面积ｎ（或背景面积ｍ）即可求得ＰＵ。公式如下：ＰＵ＝１００×｜Ｇα－ＧＡ｜／（ＡＡβ·Ｇｍａｘ）＝１００×｜Ｇα－ＧＡ｜／［（１－ＡＡα）·ｍａｘ］。式中ＡＡα＝ｎ／（ｍ＋ｎ），ＡＡβ＝ｍ／（ｍ＋ｎ），Ｇｍａｘ为测试仪器的最大灰度分级。本法灰度测试无需考虑灰度设定方法，只需在同一灰度设定条件下测试即可。本法还可用于其它组织化学、细胞化学和原位分子杂交反应显色程度定量。

An advanced quantitative method of immunohistochernical staining was developed,which brought about a new formula for testing and calculating positive unit(PU).According to this method,only the gray level and the area of defined area A and the phase a of positive products(or background 9) in A need to be tested; besides,the gray setting method needn't to be regarded.The formula is as follows:PU=100×|Ga-GA |/(AAβ.Gmax)=100×|Ga-GA|/[(1 -AAα).Gmax].G.and GA denote separately the mean gray level of the studying structure a and the test area A which equals area a plus area β in the formula;AAαand AAα mean respectively the area density of a and 5 in the A; Gmax is the maximum gray level of the instrument.Although the gray setting is free,the test of gray level should be performed in the same setting.The method could also be used for the reactive intensity of other histochemistry.cytochemistry and hybridization in situ.