Person:

Jiang, Zhe

Recording intracellular bioelectrical signals is central to understanding the fundamental behaviour of cells and cell-networks in, for example, neural and cardiac systems1–4. The standard tool for intracellular recording, the patch-clamp micropipette5 is widely applied, yet remains limited in terms of reducing the tip size, the ability to reuse the pipette5, and ion exchange with the cytoplasm6. Recent efforts have been directed towards developing new chip-based tools1–4,7–13, including micro-to-nanoscale metal pillars7–9, transistor-based kinked nanowire10,11 and nanotube devices12,13. These nanoscale tools are interesting with respect to chip-based multiplexing, but, to date, preclude targeted recording from specific cell regions and/or subcellular structures. Here we overcome this limitation in a general manner by fabricating free-standing probes where a kinked silicon nanowire with encoded field-effect transistor detector serves as the tip end. These probes can be manipulated in three dimensions (3D) within a standard microscope to target specific cells/cell regions, and record stable full-amplitude intracellular action potentials from different targeted cells without the need to clean or change the tip. Simultaneous measurements from the same cell made with free-standing nanowire and patch-clamp probes show that the same action potential amplitude and temporal properties are recorded without corrections to the raw nanowire signal. In addition, we demonstrate real-time monitoring of changes in the action potential as different ion-channel blockers are applied to cells, and multiplexed recording from cells by independent manipulation of two free-standing nanowire probes.

The miniaturization of bioelectronic intracellular probes with a wide dynamic frequency range can open up opportunities to study biological structures inaccessible by existing methods in a minimally invasive manner. Here, we report the design, fabrication, and demonstration of intracellular bioelectronic devices with probe sizes less than 10 nm. The devices are based on a nanowire–nanotube heterostructure in which a nanowire field-effect transistor detector is synthetically integrated with a nanotube cellular probe. Sub-10-nm nanotube probes were realized by a two-step selective etching approach that reduces the diameter of the nanotube free-end while maintaining a larger diameter at the nanowire detector necessary for mechanical strength and electrical sensitivity. Quasi-static water-gate measurements demonstrated selective device response to solution inside the nanotube, and pulsed measurements together with numerical simulations confirmed the capability to record fast electrophysiological signals. Systematic studies of the probe bandwidth in different ionic concentration solutions revealed the underlying mechanism governing the time response. In addition, the bandwidth effect of phospholipid coatings, which are important for intracellular recording, was investigated and modeled. The robustness of these sub-10-nm bioelectronics probes for intracellular interrogation was verified by optical imaging and recording the transmembrane resting potential of HL-1 cells. These ultrasmall bioelectronic probes enable direct detection of cellular electrical activity with highest spatial resolution achieved to date, and with further integration into larger chip arrays could provide a unique platform for ultra-high-resolution mapping of activity in neural networks and other systems.

Semiconductor nanowires and other semiconducting nanoscale materials configured as field-effect transistors have been studied extensively as biological/chemical (bio/chem) sensors. These nanomaterials have demonstrated high-sensitivity from one- and two-dimensional sensors, although the realization of the ultimate pointlike detector has not been achieved. In this regard, nanoscale p–n diodes are attractive since the device element is naturally localized near the junction, and while nanowire p–n diodes have been widely studied as photovoltaic devices, their applications as bio/chem sensors have not been explored. Here we demonstrate that p–n diode devices can serve as a new and powerful family of highly localized biosensor probes. Designed nanoscale axial p–n junctions were synthetically introduced at the joints of kinked silicon nanowires. Scanning electron microscopy images showed that the kinked nanowire structures were achieved, and electrical transport measurements exhibited rectifying behavior with well-defined turn-on in forward bias as expected for a p–n diode. In addition, scanning gate microscopy demonstrated that the most sensitive region of these nanowires was localized near the kinked region at the p–n junction. High spatial resolution sensing using these p–n diode probes was carried out in aqueous solution using fluorescent charged polystyrene nanobeads. Multiplexed electrical measurements show well-defined single-nanoparticle detection, and experiments with simultaneous confocal imaging correlate directly the motion of the nanobeads with the electrical signals recorded from the p–n devices. In addition, kinked p–n junction nanowires configured as three-dimensional probes demonstrate the capability of intracellular recording of action potentials from electrogenic cells. These p–n junction kinked nanowire devices, which represent a new way of constructing nanoscale probes with highly localized sensing regions, provide substantial opportunity in areas ranging from bio/chem sensing and nanoscale photon detection to three-dimensional recording from within living cells and tissue.

Functional kinked nanowires (KNWs) represent a new class of nanowire building blocks, in which functional devices, for example, nanoscale field-effect transistors (nanoFETs), are encoded in geometrically controlled nanowire superstructures during synthesis. The bottom-up control of both structure and function of KNWs enables construction of spatially isolated point-like nanoelectronic probes that are especially useful for monitoring biological systems where finely tuned feature size and structure are highly desired. Here we present three new types of functional KNWs including (1) the zero-degree KNW structures with two parallel heavily doped arms of U-shaped structures with a nanoFET at the tip of the “U”, (2) series multiplexed functional KNW integrating multi-nanoFETs along the arm and at the tips of V-shaped structures, and (3) parallel multiplexed KNWs integrating nanoFETs at the two tips of W-shaped structures. First, U-shaped KNWs were synthesized with separations as small as 650 nm between the parallel arms and used to fabricate three-dimensional nanoFET probes at least 3 times smaller than previous V-shaped designs. In addition, multiple nanoFETs were encoded during synthesis in one of the arms/tip of V-shaped and distinct arms/tips of W-shaped KNWs. These new multiplexed KNW structures were structurally verified by optical and electron microscopy of dopant-selective etched samples and electrically characterized using scanning gate microscopy and transport measurements. The facile design and bottom-up synthesis of these diverse functional KNWs provides a growing toolbox of building blocks for fabricating highly compact and multiplexed three-dimensional nanoprobes for applications in life sciences, including intracellular and deep tissue/cell recordings.

Semiconductor nanowire materials and devices provide unique opportunities in the frontier between nanoelectronics and biology. The bottom-up paradigm enables flexible synthesis and patterning of nanoscale building blocks with novel structures and properties, and nano-to-micro fabrication methods allow the advantages of functional nanowire elements to interface with biological systems in new ways. In this thesis, I will focus on the development of bottom-up nanoscience platforms, which includes rational synthesis and assembly of semiconductor nanowires with new capabilities, as well as design and fabrication of the first free-standing three-dimensional (3D) nanoprobes, with special focus on applications in intracellular recording and stimulation. I will first introduce kinked p-n junction nanowires as a new and powerful family of high spatial resolution biological and chemical sensors with proof-of-concept applications. Next, I will discuss a variety of functional kinked nanowires with synthetically controlled properties and the potential of achieving more detailed and less invasive cellular studies. Furthermore, I will present a general shape-controlled deterministic nanowire assembly method to produce large-scale arrays of devices with well-defined geometry and position. Then, I will present the design of a general method to fabricate these nanowire structures into free-standing 3D probes. I will show that free-standing nanowire bioprobes can be manipulated to target specific cells and record stable intracellular action potentials. I will demonstrate simultaneous measurements from the same cell using both kinked nanowire and patch-clamp probes. Moreover, I will discuss two strategies of multiplexed recording using free-standing probes. Finally, I will report localized stimulation on single cells enabled by the unique properties of p-n kinked nanowires. I will show with simulation and electrical characterization that in reverse bias, localized electric field generated around the nanoscale p-n junction should exceed the threshold for opening voltage-gated sodium channels. Moreover, I will present measurements of localized cell stimulation using p-n nanowire free-standing probes. Together with the capability of stable intracellular recording, these results complete the two-way communication between semiconductor nanowire electronics and biological systems at a natural nanoscale, which can open up new directions in the fields ranging from cellular electrophysiology, brain activity mapping to brain-machine interface.