Nanopipet Based Nanoprobes for Single-Cell Analysis

Molecular Biology has advanced our knowledge of the individual molecular components that make up living cells. The challenge, however, is to fully understand the functional integration of these components. Our group have pioneered the development of an array of new and powerful biophysical tools based on Scanning Ion Conductance Microscopy (SICM) that allow quantitative measurements and non-invasive functional imaging of the surface of living cells down to the level of single protein molecules. Scanning ion conductance microscopy and a battery of associated innovative methods are unique among current imaging techniques, not only in spatial and temporal resolution of living cells, but also in the rich combination of imaging with other functional interrogation methods [1-3]. Recently we develop a novel class of nanoprobes based on double-barrel nanopipettes. One barrel is selectively functionalized with carbon and act as a nanoelectrode while the other one is used for ion conductance microscopy. These nanoprobes allowed simultaneous electrochemical and ion conductance imaging [4] and functional mapping of the electrochemical activity of the surface of living cells [5]. Current work in our group is directed to the use of these nanoprobes for intracellular sensing. We will present recent data demonstrating their ability to measure intracellular molecules in real time both in adherent cells and in tissue.References[1] Novak et al, Neuron, 2013, 79(6), 1067-1077.[2] Novak et al, Nat Methods, 2009, 6, 279 - 281.[3] Shevchuck et al, J Cell Biol., 2012, 197(4), 499-508.[4] Takahashi et al, Angew. Chem. Int. Ed., 2011, 50(41), 9638-9642.[5] Takahashi et al, PNAS, 2012, 109 (29), 11540-11545.