Okuguchi S
Microbiol Immunol. 1978;22(3):113-21. doi: 10.1111/j.1348-0421.1978.tb00355.x.
Frauch's micro-slide method was improved to facilitate the endpoint-determination of the Limulus test. Two precise observations, by inverted phase contrast microscopy and with a staining procedure, were newly performed as additions to the slide test. The staining procedure was proposed as an improved method for the Limulus test since it is simple and convenient. In the staining method, bromophenol blue (BPB) solution was used as the staining solution. A negative (-), a strong positive (++) and a weak positive reaction (+) were characterized by a "ring" formation, a "cloud-like" spread of gel and a "spot" in the "cloud" respectively. Since the distinction between (-) and (+) reactions was obvious in the proposed method, determination of the endpoint was easier than in the ordinary tube and Frauch's method. The sensitivity of the present method was equal to or higher than that of other methods. Inverted phase contrast microscopy was utilized to confirm the findings obtained by the staining method. The volume of the lysate used in this method was as little as 1/10 of that used in the tube method.
弗劳奇微玻片法得到了改进，以促进鲎试剂检测终点的判定。在玻片检测中新增了两项精确观察，即通过倒置相差显微镜观察和采用染色程序。由于染色程序简单方便，故被提议作为鲎试剂检测的一种改进方法。在染色法中，使用溴酚蓝（BPB）溶液作为染色液。阴性（-）、强阳性（++）和弱阳性反应（+）分别以“环状”形成、凝胶的“云状”扩散以及“云”中的“斑点”为特征。由于在所提议的方法中（-）和（+）反应之间的区别明显，因此终点的判定比普通试管法和弗劳奇法更容易。本方法的灵敏度与其他方法相当或更高。利用倒置相差显微镜来确认通过染色法获得的结果。本方法中使用的裂解物体积仅为试管法所用体积的1/10。
