Direct Delivery of Nanobeads into Cells with Nanoinjector

Delivering biomolecules into living cells has become an important challenge in medical and biological fields. Conventional techniques such as virus vector and electroporation are utilized commonly for small molecule delivery, but still remain several problems of low efficiency and low viability for large molecules such as biological proteins and organelles. To overcome this problem, we demonstrate the macromolecular delivery into adhesive cells with metallic nanotube ducts, which is called as nanoinjector. The metallic nanotubes were developed by an electroless plating of gold onto track-etched polycarbonate template and wet/dry etching [1-2]. To insert Au nanotubes into the cells (such as HeLa and NIH3T3), we have developed an autostamping system that can be integrated with the microscope. Here we demonstrated the delivery of macro molecules (a few hundred nanobeads) into the cells and compare it with the endocytosis delivery. The nanoinjector system provide the high efficient delivery of 200nm diameter beads for 10min injection, while the endocytosis was at low efficiency, which takes for 24h to deliver the nanobeads. Furthermore, our nanoinjector can deliver the nanobeads uniformly into the cells.<br/>[References]<br/>[1] B. Zhang, Y. Shi, D. Miyamoto, K. Nakazawa, T. Miyake, “Nanostraw membrane stamping for direct delivery of molecules into adhesive cells”, Scientific Reports, 9, 6806, 2019.[MT1] <br/>[2] B. Zhang, D. Zheng, S. Yiming, K. Oyama, M. Ito, M. Ikari, T. Kigawa, T. Mikawa, T. Miyake, “High-Efficient and Dosage-Controllable Intracellular Cargo Delivery through Electrochemical Metal–Organic Hybrid Nanogates”, Small Sci. 1, 2100069, 2021.