Sweet G H, Schindler C A
J Bacteriol. 1967 Jun;93(6):1943-8. doi: 10.1128/jb.93.6.1943-1948.1967.
Portions of a whole antiserum to Histoplasma capsulatum were reacted with amounts of fluorescein isothiocyanate (FITC) that ranged from 50 to 400 mug/mg of protein. Portions of the globulin from the same antiserum were reacted with amounts of FITC that ranged from 12.5 to 50 mug of FITC per mg of protein. The globulin conjugates (postlabeled globulins), the whole serum conjugates, and the globulins from the whole serum conjugates (prelabeled globulins) were compared with respect to their fluorescein-protein (F:P) ratios and fluorescent-antibody (FA) activities. The whole serum sample treated with 50 mug of FITC per mg of protein was least reactive in FA tests, and its globulin had the lowest F:P. All other conjugates had globulins with F:P ratios that were considered to be adequate for high FA activity. It was found, however, that the prelabeled globulins were considerably less reactive than the postlabeled globulins or the whole serum conjugates. A larger amount of brightly staining reagent per milliliter of original serum could be obtained from labeled whole serum than from postlabeled globulin. Lissamine-rhodamine conjugated to bovine serum albumin (LRBSA) was evaluated as a counterstain to be used in conjunction with FITC-labeled whole antisera. The counterstain was effective in masking nonspecific FITC fluorescence in Formalin-fixed tissues and in culture smears of fungi. Masking was incomplete in culture smears of a bacterium and in blood smears containing a protozoan.
将针对荚膜组织胞浆菌的全抗血清的部分与异硫氰酸荧光素（FITC）进行反应，其用量范围为每毫克蛋白质50至400微克。将来自同一抗血清的球蛋白部分与每毫克蛋白质12.5至50微克的FITC进行反应。比较了球蛋白缀合物（后标记球蛋白）、全血清缀合物以及全血清缀合物中的球蛋白（预标记球蛋白）的荧光素 - 蛋白质（F:P）比率和荧光抗体（FA）活性。在FA试验中，每毫克蛋白质用50微克FITC处理的全血清样品反应性最低，其球蛋白的F:P最低。所有其他缀合物的球蛋白F:P比率被认为足以产生高FA活性。然而，发现预标记球蛋白的反应性明显低于后标记球蛋白或全血清缀合物。与后标记球蛋白相比，从标记的全血清中每毫升原始血清可获得更多量的亮染色试剂。评估了与牛血清白蛋白缀合的丽丝胺罗丹明（LRBSA）作为与FITC标记的全抗血清联合使用的复染剂。该复染剂可有效掩盖福尔马林固定组织和真菌培养涂片中非特异性FITC荧光。在细菌培养涂片和含有原生动物的血涂片中，掩盖不完全。
