An equipment for microdetermination of superoxide production of nentrophils.

我们开发了一种用于多形核中性粒细胞（PMNs）超氧化物生成的微量测定设备。该设备由专用双光束分光光度计（550nm - 540nm）和微机数据分析系统组成，用于消除样品中细胞产生的噪声。肉豆酸酯或豆豆蛋白A +细胞charasin D （FMLP）刺激PMNs，对富含PMNs的血浆的最佳刺激量为0.01 ~ 5 ug，对经PMA分离的PMNs的最佳刺激量为0.1 ~ 5 ug；Con A加Cyt D为10 ~ 100 ug， FMLP为0.1 ~ 1 mol。单次测定仅需5 ~ 10x10^4个PMNs。通过PMA分析，富等离子体PMNs的超氧化物生成速率与分离PMNs的超氧化物生成速率具有良好的相关性。通过Con A + Cyt D检测，新生儿血浆富集pmn值与分离pmn值之间的超氧化物生成率呈良好的相关性，而成人血浆富集pmn值低于分离pmn值。这种新装置为我们方便地测定pmn的超氧化物生成速率提供了条件，并且可以利用新生儿血浆富集pmn来测定超氧化物生成速率。

We have developped anequipment for mictrodetermination of superoxide production of polymorphonuclear neutrophils (PMNs).This equipment consists of special double beam spectrophotometer (550nm - 540nm) and data analysis system by microcomputer for eliminating a noise due to cells in the samples.PMNs were stimulated by either phorbol myristate acetate or Concanavalin A plus Cytocharasin D, Or FMLP, and their optimal Quantities were 0.01 to 5 ug for PMNs riched plasma, 0.1 to 5 ug for separated PMNs by PMA; 10 to 100 ug for Con A plus Cyt D, 0.1 to 1 umoles for FMLP. Only 5 to 10x10^4 PMNs were required for single/assay.By PMA, the superoxide production rates showed a good correlation between the value of plasma riched PMNs and the value of separated PMNs. By Con A plus Cyt D, The superoxide production rates showed a good correlation between the value of plasma riched PMNs and the values of separated PMNs in Newborns, however in adult, the values obtained from plasma riched PMNs were lower than those obtained from separated PMNs.This new device provided us to determine superoxide production rates of PMNs conveniently, and it is possible to determine it using plasma riched PMNs in newborn infants.