Microneedle Patch for Painless Intradermal Collection of Interstitial Fluid Enabling Multianalyte Measurement of Small Molecules, SARS-CoV-2 Antibodies, and Protein Profiling.

Blood sampling is a common practice to monitor health, but it entails a series of drawbacks for patients including pain and discomfort. Thus, there is a demand for more convenient ways to obtain samples. Modern analytical techniques enable monitoring of multiple bioanalytes in smaller samples, opening possibilities for new matrices, and microsampling technologies to be adopted. Interstitial fluid (ISF) is an attractive alternative matrix that shows good correlation with plasma concentration dynamics for several analytes and can be sampled in a minimally invasive and painless manner from the skin at the point-of-care. However, there is currently a lack of sampling devices compatible with clinical translation. Here, to tackle state-of-the-art limitations, a cost-effective and compact single-microneedle-based device designed to painlessly collect precisely 1.1 µL of dermal ISF within minutes is presented. The fluid is volume-metered, dried, and stably stored into analytical-grade paper within the microfluidic device. The obtained sample can be mailed to a laboratory, quantitatively analyzed, and provide molecular insights comparable to blood testing. In a human study, the possibility to monitor various classes of molecular analytes is demonstrated in ISF microsamples, including caffeine, hundreds of proteins, and SARS-CoV-2 antibodies, some being detected in ISF for the first time.

Vertical integration of microchips by magnetic assembly and edge wire bonding.

The out-of-plane integration of microfabricated planar microchips into functional three-dimensional (3D) devices is a challenge in various emerging MEMS applications such as advanced biosensors and flow sensors. However, no conventional approach currently provides a versatile solution to vertically assemble sensitive or fragile microchips into a separate receiving substrate and to create electrical connections. In this study, we present a method to realize vertical magnetic-field-assisted assembly of discrete silicon microchips into a target receiving substrate and subsequent electrical contacting of the microchips by edge wire bonding, to create interconnections between the receiving substrate and the vertically oriented microchips. Vertical assembly is achieved by combining carefully designed microchip geometries for shape matching and striped patterns of the ferromagnetic material (nickel) on the backside of the microchips, enabling controlled vertical lifting directionality independently of the microchip's aspect ratio. To form electrical connections between the receiving substrate and a vertically assembled microchip, featuring standard metallic contact electrodes only on its frontside, an edge wire bonding process was developed to realize ball bonds on the top sidewall of the vertically placed microchip. The top sidewall features silicon trenches in correspondence to the frontside electrodes, which induce deformation of the free air balls and result in both mechanical ball bond fixation and around-the-edge metallic connections. The edge wire bonds are realized at room temperature and show minimal contact resistance (<0.2 Ω) and excellent mechanical robustness (>168 mN in pull tests). In our approach, the microchips and the receiving substrate are independently manufactured using standard silicon micromachining processes and materials, with a subsequent heterogeneous integration of the components. Thus, this integration technology potentially enables emerging MEMS applications that require 3D out-of-plane assembly of microchips.

Real-time intradermal continuous glucose monitoring using a minimally invasive microneedle-based system.

Continuous glucose monitoring (CGM) has the potential to greatly improve diabetes management. The aim of this work is to show a proof-of-concept CGM device which performs minimally invasive and minimally delayed in-situ glucose sensing in the dermal interstitial fluid, combining the advantages of microneedle-based and commercially available CGM systems. The device is based on the integration of an ultra-miniaturized electrochemical sensing probe in the lumen of a single hollow microneedle, separately realized using standard silicon microfabrication methods. By placing the sensing electrodes inside the lumen facing an opening towards the dermal space, real-time measurement purely can be performed relying on molecular diffusion over a short distance. Furthermore, the device relies only on passive capillary lumen filling without the need for complex fluid extraction mechanisms. Importantly, the transdermal portion of the device is 50 times smaller than that of commercial products. This allows access to the dermis and simultaneously reduces tissue trauma, along with being virtually painless during insertion. The three-electrode enzymatic sensor alone was previously proven to have satisfactory sensitivity (1.5 nA/mM), linearity (up to 14 mM), selectivity, and long-term stability (up to 4 days) in-vitro. In this work we combine this sensor technology with microneedles for reliable insertion in forearm skin. In-vivo human tests showed the possibility to correctly and dynamically track glycaemia over time, with approximately 10 min delay with respect to capillary blood control values, in line with the expected physiological lag time. The proposed device can thus reduce discomfort and potentially enable less invasive real-time CGM in diabetic patients.

Ultra-miniaturization of a planar amperometric sensor targeting continuous intradermal glucose monitoring.

An ultra-miniaturized electrochemical biosensor for continuous glucose monitoring (CGM) is presented. The aim of this work is to demonstrate the possibility of an overall reduction in sensor size to allow minimally invasive glucose monitoring in the interstitial fluid in the dermal region, in contrast to larger state-of-the-art systems, which are necessarily placed in the subcutaneous layer. Moreover, the reduction in size might be a key factor to improve the stability and reliability of transdermal sensors, due to the reduction of the detrimental foreign body reaction and of consequent potential failures. These advantages are combined with lower invasiveness and discomfort for patients. The realized device consists of a microfabricated three-electrode enzymatic sensor with a total surface area of the sensing portion of less than 0.04mm2, making it the smallest fully integrated planar amperometric glucose sensor area reported to date. The working electrode and counter electrode consist of platinum and are functionalized by drop casting of three polymeric membranes. The on-chip iridium oxide (IrOx) pseudo-reference electrode provides the required stability for measurements under physiological conditions. The device is able to dynamically and linearly measure glucose concentrations in-vitro over the relevant physiological range, while showing sufficient selectivity to known interfering species present in the interstitial fluid, with resolution and sensitivity (1.51nA/mM) comparable to that of state-of-art commercial CGM systems. This work can therefore enable less invasive and improved CGM in patients affected by diabetes.