Pepperkok R, Schneider C, Philipson L, Ansorge W
European Molecular Biology Laboratory, Heidelberg, FRG.
Cytotechnology. 1991;5 Suppl 1:93-8.
An automated capillary microinjection system with computer controlled positioning of the cells and of the capillary, and its applications and advantages are described. About 1500 injections are possible in one hour, with high reproducibility. In cytoplasmic and nuclear injections more than 90% and 85% of the cells are successfully injected. Using FITC-Dextran at a concentration of 0.5% as a fluorescently labelled coinjection marker, 99% of the cells can be retrieved in culture medium even 48 hours after injection. The coordinates of the cells are stored in the computer and accuracy in statistical evaluation of experiments is improved in comparison to the manual techniques. Methods for preparation and handling of glass capillaries were developed resulting in reproducible form and significantly reduced clogging rate. The improved characteristics offered by this system are demonstrated in studies leading to the confirmation of existence of mRNA(s) inhibiting cell proliferation. Functional screening by cell injections of cDNA libraries and of size fractionated mRNA molecules can be performed efficiently with the automated microinjection system.
描述了一种具有计算机控制细胞和毛细管定位功能的自动毛细管显微注射系统及其应用和优点。一小时内大约可以进行1500次注射，重现性高。在细胞质和细胞核注射中，分别有超过90%和85%的细胞被成功注射。使用浓度为0.5%的异硫氰酸荧光素标记葡聚糖作为荧光共注射标记物，即使在注射48小时后，99%的细胞仍可在培养基中回收。细胞坐标存储在计算机中，与手动技术相比，实验统计评估的准确性得到提高。开发了玻璃毛细管的制备和处理方法，使其形状可重现，堵塞率显著降低。该系统提供的改进特性在导致证实存在抑制细胞增殖的mRNA的研究中得到了证明。利用自动显微注射系统可以高效地通过细胞注射对cDNA文库和大小分级的mRNA分子进行功能筛选。
