Mechanical sensing of the penetration of various nanoneedles into a living cell using atomic force microscopy

Obataya, Ikuo; Nakamura, Chikashi; Han, Shiyu; Nakamura, Noriyuki; Miyake, Jun

doi:10.1016/j.bios.2004.07.020

Cited by 117 publications

(121 citation statements)

References 12 publications

(15 reference statements)

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“…2, and Supplementary Note 1). These results are consistent with a previous report by Obataya et al 23 , which suggested that a vertical sidewall is more suitable for rapid penetration of cell membrane.…”

Section: Resultssupporting

confidence: 83%

Poking cells for efficient vector-free intracellular delivery

et al. 2014

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Techniques for introducing foreign molecules and materials into living cells are of great value in cell biology research. A major barrier for intracellular delivery is to cross the cell membrane. Here we demonstrate a novel platform utilizing diamond nanoneedle arrays to facilitate efficient vector-free cytosolic delivery. Using our technique, cellular membrane is deformed by an array of nanoneedles with a force on the order of a few nanonewtons. We show that this technique is applicable to deliver a broad range of molecules and materials into different types of cells, including primary neurons in adherent culture. Especially, for delivering plasmid DNAs into neurons, our technique produces at least eightfold improvement (B45% versus B1-5%) in transfection efficiency with a dramatically shorter experimental protocol, when compared with the commonly used lipofection approach. It is anticipated that our technique will greatly benefit basic research in cell biology and also a wide variety of clinical applications.

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Section: Resultssupporting

confidence: 83%

Poking cells for efficient vector-free intracellular delivery

et al. 2014

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“…There is a strong desire among researchers to directly observe molecular processes occurring in the cell without using optical markers, raising the question of whether it is possible to look at the cell interior with HS-AFM. It has already been demonstrated that the long tip (even a thin pipette) can be inserted into the cell without killing the cell (Obataya et al 2005;Guillaume-Gentil et al 2014). However, no one has tried to carry out AFM imaging of the cell interior.…”

Section: Nano-endoscopymentioning

confidence: 99%

High-speed atomic force microscopy and its future prospects

2017

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Various techniques have been developed and used to investigate how proteins produce complex biological architectures and phenomena. Among these techniques, high-speed atomic force microscopy (HS-AFM) holds a unique position. It is only HS-AFM that allows the simultaneous assessment of structure and dynamics of single protein molecules in action. This new microscopy tool has been successfully applied to a variety of proteins, from motor proteins to membrane proteins, antibodies, enzymes, and even to intrinsically disordered proteins. And yet there still remain many biomolecular phenomena that cannot be addressed by HS-AFM in its current form. Here, I present a brief history of HS-AFM development, describe the current state of HS-AFM, and then discuss which new biological scanning probe microscopy techniques will be coming up next.

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“…To this end, glass microneedles are pushed through the membrane into the nucleus by means of a micromanipulator. Recently, a modified AFM using nanoneedles was developed (Obataya et al 2005a). These nanoneedles can be used to penetrate the cell membrane to manipulate internal cellular structures (Obataya et al 2005b).…”

Section: Intracellular Manipulationmentioning

confidence: 99%

Intracellular manipulation of chromatin using magnetic nanoparticles

2008

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Magnetic tweezers are widely used for manipulating small magnetic beads inside the cell cytoplasm in order to gain insight into the structural and mechanical properties of the cytoskeleton. Here we discuss the use of magnetic tweezers for the study of nuclear architecture and the mechanical properties of chromatin in living cells. A custom-built, dedicated micro magnetic tweezer set-up is described. We review progress that has been made in applying this technology for the study of chromatin structure and discuss its prospects for the in situ analysis of nuclear architecture and chromatin function.

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Mechanical sensing of the penetration of various nanoneedles into a living cell using atomic force microscopy

Cited by 117 publications

References 12 publications

Poking cells for efficient vector-free intracellular delivery

Poking cells for efficient vector-free intracellular delivery

High-speed atomic force microscopy and its future prospects

Intracellular manipulation of chromatin using magnetic nanoparticles

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