Matsumoto Daisuke, Yamagishi Ayana, Saito Megumi, Sathuluri Ramachandra Rao, Silberberg Yaron R, Iwata Futoshi, Kobayashi Takeshi, Nakamura Chikashi
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 5 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
J Biosci Bioeng. 2016 Dec;122(6):748-752. doi: 10.1016/j.jbiosc.2016.05.006. Epub 2016 Jun 15.
Efficient and rapid delivery of macromolecule probes, such as quenchbodies and other large biomarkers that cannot readily pass through the plasma membrane, is necessary for live-cell imaging and other intracellular analyses. We present here an alternative, simple method for delivery of macromolecules into live cells. In this method, which we term here mechanoporation, a nanoneedle array is used for making transient pores in the plasma membrane to allow access of desired macromolecules into thousands of live cells, simultaneously. This rapid, 3-step method facilitates an efficient delivery by adding macromolecules into the medium, inserting nanoneedles into the cells and oscillating the nanoneedle array, a process that takes no more than 5 min in total. In addition, we demonstrate here how this method can repeatedly and reproducibly deliver molecules into specifically-selected locations on a given cell culture dish. The results presented here show how this unique mechanoporation method enables rapid and high-throughput bio-macromolecule delivery and live-cell imaging.
对于活细胞成像和其他细胞内分析而言，高效快速地递送大分子探针（如猝灭体和其他难以轻易穿过质膜的大型生物标志物）是必要的。我们在此介绍一种将大分子递送至活细胞的简单替代方法。在我们称为机械穿孔的此方法中，使用纳米针阵列在质膜上制造瞬时孔，以使所需大分子能够同时进入数千个活细胞。这种快速的三步法通过将大分子添加到培养基中、将纳米针插入细胞并振荡纳米针阵列来促进高效递送，整个过程总共不超过5分钟。此外，我们在此展示了该方法如何能够反复且可重复地将分子递送至给定细胞培养皿上的特定选定位置。此处呈现的结果表明了这种独特的机械穿孔方法如何实现快速且高通量的生物大分子递送和活细胞成像。
