Scalable Fabrication Framework of Implantable Ultrathin and Flexible Probes with Biodegradable Sacrificial Layers

For long-term biocompatibility and performance, implanted probes need to further reduce their size and mechanical stiffness to match that of the surrounding cells, which, however, makes accurate and minimally invasive insertion operations difficult due to lack of rigidity and brings additional complications in assembling and surgery. Here, we report a scalable fabrication framework of implantable probes utilizing biodegradable sacrificial layers to address this challenge. Briefly, the integrated biodegradable sacrificial layer can dissolve in physiological fluids shortly after implantation, which allows the in situ formation of functional ultrathin film structures off of the initial small and rigid supporting backbone. We show that the dissolution of this layer does not affect the viability and excitability of neuron cells in vitro. We have demonstrated two types of probes that can be used out of the box, including (1) a compact probe that spontaneously forms three-dimensional bend-up devices only after implantation and (2) an ultraflexible probe as thin as 2 μm attached to a small silicon shaft that can be accurately delivered into the tissue and then get fully released in situ without altering its shape and position because the support is fully retracted. We have obtained a >93% yield of the bend-up structure, and its geometry and stiffness can be systematically tuned. The robustness of the ultraflexible probe has been tested in tissue-mimicking agarose gels with <1% fluctuation in the test resistance. Our work provides a general strategy to prepare ultrasmall and flexible implantable probes that allow high insertion accuracy and minimal surgical damages with the best biocompatibility

Multiplexed Free-Standing Nanowire Transistor Bioprobe for Intracellular Recording: A General Fabrication Strategy

Recent advance in free-standing nanowire transistor bioprobes opens up new opportunities of accurately interfacing spatially unobstructed nanoscale sensors with live cells. However, the existing fabrication procedures face efficiency and yield limitations when working with more complex nanoscale building blocks to integrate, for example, multiplexed recordings or additional functionalities. To date, only single-kinked silicon nanowires have been successfully used in such probes. Here we establish a general fabrication strategy to mitigate such limitations with which synthetically designed complex nanoscale building blocks can be readily used without causing significant penalty in yield or fabrication time, and the geometry of the probe can be freely optimized based on the orientation and structure of the building blocks. Using this new fabrication framework, we demonstrate the first multiplexed free-standing bioprobe based on w-shaped silicon kinked nanowires that are synthetically integrated with two nanoscale field-effect transistor devices. Simultaneous recording of intracellular action potentials from both devices have been obtained of a single spontaneously beating cardiomyocyte

Design and Synthesis of Diverse Functional Kinked Nanowire Structures for Nanoelectronic Bioprobes

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

Synthetically Encoded Ultrashort-Channel Nanowire Transistors for Fast, Pointlike Cellular Signal Detection

Nanostructures, which have sizes comparable to biological functional units involved in cellular communication, offer the potential for enhanced sensitivity and spatial resolution compared to planar metal and semiconductor structures. Silicon nanowire (SiNW) field-effect transistors (FETs) have been used as a platform for biomolecular sensors, which maintain excellent signal-to-noise ratios while operating on lengths scales that enable efficient extra- and intracellular integration with living cells. Although the NWs are tens of nanometers in diameter, the active region of the NW FET devices typically spans micrometers, limiting both the length and time scales of detection achievable with these nanodevices. Here, we report a new synthetic method that combines gold-nanocluster-catalyzed vapor–liquid–solid (VLS) and vapor–solid–solid (VSS) NW growth modes to produce synthetically encoded NW devices with ultrasharp (<5 nm) n-type highly doped (n⁺⁺) to lightly doped (n) transitions along the NW growth direction, where n⁺⁺ regions serve as source/drain (S/D) electrodes and the n-region functions as an active FET channel. Using this method, we synthesized short-channel n⁺⁺/n/n⁺⁺ SiNW FET devices with independently controllable diameters and channel lengths. SiNW devices with channel lengths of 50, 80, and 150 nm interfaced with spontaneously beating cardiomyocytes exhibited well-defined extracellular field potential signals with signal-to-noise values of ca. 4 independent of device size. Significantly, these “pointlike” devices yield peak widths of ∼500 μs, which is comparable to the reported time constant for individual sodium ion channels. Multiple FET devices with device separations smaller than 2 μm were also encoded on single SiNWs, thus enabling multiplexed recording from single cells and cell networks with device-to-device time resolution on the order of a few microseconds. These short-channel SiNW FET devices provide a new opportunity to create nanoscale biomolecular sensors that operate on the length and time scales previously inaccessible by other techniques but necessary to investigate fundamental, subcellular biological processes

Outside Looking In: Nanotube Transistor Intracellular Sensors

Nanowire-based field-effect transistors, including devices with planar and three-dimensional configurations, are being actively explored as detectors for extra- and intracellular recording due to their small size and high sensitivities. Here we report the synthesis, fabrication, and characterization of a new needle-shaped nanoprobe based on an active silicon nanotube transistor, ANTT, that enables high-resolution intracellular recording. In the ANTT probe, the source/drain contacts to the silicon nanotube are fabricated on one end, passivated from external solution, and then time-dependent changes in potential can be recorded from the opposite nanotube end via the solution filling the tube. Measurements of conductance versus water-gate potential in aqueous solution show that the ANTT probe is selectively gated by potential changes within the nanotube, thus demonstrating the basic operating principle of the ANTT device. Studies interfacing the ANTT probe with spontaneously beating cardiomyocytes yielded stable intracellular action potentials similar to those reported by other electrophysiological techniques. In addition, the straightforward fabrication of ANTT devices was exploited to prepare multiple ANTT structures at the end of single probes, which enabled multiplexed recording of intracellular action potentials from single cells and multiplexed arrays of single ANTT device probes. These studies open up unique opportunities for multisite recordings from individual cells through cellular networks

Fixed-Gap Tunnel Junction for Reading DNA Nucleotides

Previous measurements of the electronic conductance of DNA nucleotides or amino acids have used tunnel junctions in which the gap is mechanically adjusted, such as scanning tunneling microscopes or mechanically controllable break junctions. Fixed-junction devices have, at best, detected the passage of whole DNA molecules without yielding chemical information. Here, we report on a layered tunnel junction in which the tunnel gap is defined by a dielectric layer, deposited by atomic layer deposition. Reactive ion etching is used to drill a hole through the layers so that the tunnel junction can be exposed to molecules in solution. When the metal electrodes are functionalized with recognition molecules that capture DNA nucleotides <i>via</i> hydrogen bonds, the identities of the individual nucleotides are revealed by characteristic features of the fluctuating tunnel current associated with single-molecule binding events