激光电泳测量条件及影响因素

The Influencing Factors in Measuring Electrophoretic Mobility by Laser Doppler Spectroscopy

研究激光电泳测量条件及影响因素发现：在实体瘤制备单细胞过程中，胶原酶消化获得的单细胞其电泳迁移率高于机械消化法，但在正常组织中胶原酶消化与机械消化法所得细胞的电泳迁移率并无差异。因此，消化酶的使用影响细胞表面电荷的测定。在细胞预处理过程中，醛化与未醛化细胞其电泳迁移率一致，醛化不影响细胞表面电荷的测定。改变细胞浓度，其电泳迁移率随浓度的升高而下降，红细胞浓度＞２×１０７／ｍｌ，其电泳迁移率存在一饱和值，肝素抗凝影响红细胞的电泳迁移率。因此，对激光电泳生物定标选择不加肝素抗凝的人全血，细胞浓度为３×１０７／ｍｌ。在２４℃、３７℃测量细胞电泳迁移率无改变，故激光电泳在此温度范围内其电泳迁移率具有温度稳定性。

This study endeavored to identify the factors that influence the measurement of electrophoretic mobility(EPM)of singled cells using Laser Doppler Spectroscopy.The cells of solid turmors prepared with collagenase digestion had a higher EPM than those tumor cells prepared mechanically.No such disparity was observed in normal tissue cells.This indicated that in the process of preparation of singled cells the use of digestive enzyme affected cell surface electric charge.Fixation of cell with 0.025% plata-di-aldehyde did not alter EPM,hence no effect on cell surface charges.Alteration of concertration of singled cells was another factor that affected the EMP,which declined with the increase of cellular concentration.In measuring EPM of red blood cells, the addition of anticoagulant,heparin affected the EPM. No variation of the EPM was observed under temperature range of 24℃to 37℃.