细胞电穿孔动态过程的荧光测量

利用改进后的Ｔｈ／ＤＰＡ荧光方法及探针ＥＢ对人血影及大鼠骨髓细胞电穿孔的动态过程及其与电脉冲参数的关系进行了系统的研究．测量结果表明，在临界点以上电场作用下，血影电穿孔在电击后０．２—０．３ｓ时达最大，在约０．８ｓ时愈合；而大鼠骨髓细胞电穿孔在电击后０．４—０．９ｓ达到最大，３－５ｓ左右愈合；电穿孔大小及扩大、愈合速率与电脉冲参数有关。１０－４０ｍｍｏｌ／Ｌ乙醇和５－２０ｍｍｏｌ／Ｌ成二醛抑制血影对Ｔｂ３＋离子的电通透，相同浓度的成二醛作用强于乙醇。这些结果将为电穿孔技术的合理应用提供参考。

DYNAMIC STUDIES ON ELECTROPORATION OF CELL MEMBRANE BY FLUOROMETRIC METHOD

ifferent electric pulses with duration of 10-50us and strength of 1.0-3.0kV/cm were ap plied to suspended human erythrocyte ghosts loaded with Tb3 and the changes of fluorescence with the were immediately measured yb modified Tb/DP A method.The experimental results show that: under certain condition the electropore was formed and its area increased rapidly and then decreased. The total electropore area reached a maximum within 200-300ms and then resealed within about 800m. The electropore area and the rate of pore enlarging and resealing were related to the combination of the electric pulse palameters and other factors which affect the membrane. The inhibition of 10-40mmol/L ethanol or 5-20mmol/L glutalaldehyde. on ghost electroporation was detected and glutaralde-hyde showed stronger effect than ethanol under the same condition. Similar results were obtained in rat bone marrow cells suspended in 25 ug/ml EB, exaept for the longer pore enarging and resealing time which are 0.4-0.9s and 3-5s respectively.These results will will provide usedful information for the electropoaration techniques,especially for the cell electrofusion and the gene electroinjection.