European Molecular Biology Laboratory, Heidelberg, Germany.
Cytotechnology. 1991 Feb;5(Suppl 1):93-8. doi: 10.1007/BF00736821.
An automated capillary microinjection system with computer controlled positioning of the cells and of the capillary, and its applications and advantages arc 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 cultrue medium even 48 hours after unjection. The coordinates of the cells are stored in the computer and accuracy in statistical evaluation of experiments is improved in coparison to the manual techniques. Methods for preparatin 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.
描述了一种带有计算机控制的细胞和毛细管定位的自动化毛细管微量注射系统及其应用和优点。大约每小时可以进行 1500 次注射，具有很高的重现性。在细胞质和核内注射中，超过 90%和 85%的细胞成功注射。使用浓度为 0.5%的 FITC-Dextran 作为荧光标记的共注射标记物，即使在注射后 48 小时，也可以在培养介质中回收 99%的细胞。细胞的坐标存储在计算机中，与手动技术相比，实验的统计评估的准确性得到了提高。开发了用于制备和处理玻璃毛细管的方法，从而使毛细管具有可重复的形状和显著降低的堵塞率。通过细胞注射 cDNA 文库和大小分级的 mRNA 分子进行的功能筛选可以使用自动化微量注射系统有效地进行。
