Mechanoporation of living cells for delivery of macromolecules using nanoneedle array.
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.
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
Daisuke
D
Matsumoto
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 5 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
Ayana
A
Yamagishi
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
Megumi
M
Saito
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 5 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
Ramachandra Rao
RR
Sathuluri
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 5 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
Yaron R
YR
Silberberg
Department of Mechanical Engineering, Shizuoka University, 3-5-1 Johoku, Hamamatsu, Shizuoka 432-8561, Japan.
Futoshi
F
Iwata
Research Center for Ubiquitous MEMS and Micro Engineering, AIST, 1-2-1 Namiki, Tsukuba, Ibaraki 305-8564, Japan.
Takeshi
T
Kobayashi
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. Electronic address: chikashi-nakamura@aist.go.jp.
Chikashi
C
Nakamura
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002463:Cell Membrane Permeability; D058948:Cell Tracking / Q000379:methods*; D003911:Dextrans / Q000493:pharmacokinetics; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D057809:HEK293 Cells; D006801:Humans; D046911:Macromolecular Substances / Q000493:pharmacokinetics*; D051379:Mice; D041681:NIH 3T3 Cells; D049329:Nanostructures; D009339:Needles; D059010:Single-Cell Analysis / Q000379:methods
D003911:Dextrans; D046911:Macromolecular Substances; C015219:fluorescein isothiocyanate dextran; D016650:Fluorescein-5-isothiocyanate
Continuous and rapid delivery; Mammalian cells; Mechanoporation; Nanoneedle array; Position-specific delivery