The widespread adaptation of 3D printing in the microfluidic, bioelectronic, and Bio-MEMS communities has been stifled by the lack of investigation into the biocompatibility of commercially available printer resins. By introducing an in-depth post-printing treatment of these resins, their biocompatibility can be dramatically improved up to that of a standard cell culture vessel (99.99%). Additionally, encapsulating resins that are less biocompatible with materials that are common constituents in biosensors further enhances the biocompatibility of the material. This investigation provides a clear pathway toward developing fully functional and biocompatible 3D printed biosensor devices, especially for interfacing with electrogenic cells, utilizing benchtop-based microfabrication, and post-processing techniques.

Laser micromachining is a direct write microfabrication technology that has several advantages over traditional micro/nanofabrication techniques. In this paper, we present a comprehensive characterization of a QuikLaze 50ST2 multimodal laser micromachining tool by determining the ablation characteristics of six (6) different materials and demonstrating two applications. Both the thermodynamic theoretical and experimental ablation characteristics of stainless steel (SS) and aluminum are examined at 1064 nm, silicon and polydimethylsiloxane (PDMS) at 532 nm, and Kapton^® and polyethylene terephthalate at 355 nm. We found that the experimental data aligned well with the theoretical analysis. Additionally, two applications of this multimodal laser micromachining technology are demonstrated: shadow masking down to approximately 1.5 µm feature sizes and interdigitated electrode (IDE) fabrication down to 7 µm electrode gap width.

Microelectrode Arrays are established platforms for biosensing applications; however, limitations in electrode impedance and cell-electrode coupling still exist. In this paper, the SNR of 25 μm diameter gold (Au) microelectrodes was improved by decreasing the impedance with precision electrodeposition. SEM determined that N-P Pt. microelectrodes had nano-porous structures that filled the insulation cylinders. EIS, CV, and RMS noise measurements concluded that the optimized electrodeposition of N-P Pt. led to a lowered impedance of 18.36 kΩ ± 2.6 kΩ at 1 kHz, a larger double layer capacitance of 73 nF, and lowered RMS noise of 2.08 ± 0.16 μV as compared to the values for Au of 159 kΩ ± 28 kΩ at 1 kHz, 17nF, and 3.14 ± 0.42 μ V, respectively. Human motoneurons and human cardiomyocytes were cultured on N-P Pt. devices to assess their biocompatibility and signal quality. In order to improve the cell-electrode coupling, a precision plating technique was used. Both cell types were electrically active on devices for up to 10 weeks, demonstrated improved SNR, and expected responses to precision chemical and electrical stimulation. The modification of Au microelectrodes with nanomaterials in combination with precision culturing of human cell types provides cost effective, highly sensitive, well coupled and relevant biosensing platforms for medical and pharmaceutical research. [2018-0187]

Microfabrication and assembly of 3D MEA based on a glass-stainless steel platform is shown utilizing non-traditional “Makerspace Microfabrication” techniques featuring cost-effective, rapid fabrication and an assorted biocompatible material palette.

Adverse cardiac events are a major cause of late-stage drug development withdrawals. Improved in vitro systems for predicting cardiotoxicity are of great interest to prevent these events and to reduce the expenses involved in the introduction of cardiac drugs into the marketplace. Interdigitated electrodes (IDEs) affixed with a culture well provide a simple, suitable solution for in vitro analysis of cells because of their high sensitivity, ease of fabrication, and label-free, nondestructive analysis. Culturing human pluripotent stem cell differentiated cardiomyocytes onto these IDEs allows for the use of the IDE–cell combination in predictive toxicity assays. IDEs with smaller interdigitated distances allow for greater sensitivity, but typically require cleanroom fabrication. In this communication, we report the definition of a simple IDE geometry on a printed nanostructured substrate, demonstrate a Cellular Index (CI) increase from 0 to 7.7 for human cardiomyocytes, and a decrease in CI from 2.3 to 1 with increased concentration of the model drug, norepinephrine. The nanostructuring results in an increased sensitivity of our 1 mm pitch IDEs when compared to traditionally fabricated IDEs with a pitch of 10 μm (100 times larger electrode gap). The entire nanostructured IDE (nIDE) is fabricated and assembled in a rapid nanofabrication environment, thus allowing for iterative design changes and robust fabrication of devices.

In this work, preliminary investigations into the different sources of sensitivity enhancement of Interdigitated Electrodes (IDEs) on polymer substrates utilized in cell-based biosensing are evaluated. In studying underlying modalities involved in this IDE sensitivity enhancement, we have found that this phenomenon has several components - geometric (electrode gap width), nanostructuring of the substrate, and surface plasmon resonance (SPR) enhancement modes. By isolating these three modes, this article investigates and reports on the individual contribution of each of these modes in enhancing the sensitivity of IDEs during a short five days in vitro (5 DIV) assay of the HL-1 cardiomyocytes cultured on these devices. [2020-0121]