Hyeonaug Hong

Three-Dimensional Rapid Prototyping of Multidirectional Polymer Nanoprobes for Single Cell Insertion

Three-dimensional (3D) thermal drawing at nanoscale as a novel rapid prototyping method was demonstrated to create multidirectional polymer nanoprobes for single cell analysis. This 3D drawing enables simple and rapid fabrication of polymeric nanostructures with high aspect ratio. The effect of thermal drawing parameters, such as drawing speeds, dipping depths, and contact duration on the final geometry of polymer nanostructures was investigated. Vertically aligned and L-shaped nanoprobes were fabricated and their insertion into living single cells such as algal cells and human neural stem cells was demonstrated. This technique can be extended to create more complex 3D structures by controlling drawing steps and directions on any surface.

Insertion of Vertically Aligned Nanowires into Living Cells by Inkjet Printing of Cells

Effective insertion of vertically aligned nanowires (NWs) into cells is critical for bioelectrical and biochemical devices, biological delivery systems, and photosynthetic bioenergy harvesting. However, accurate insertion of NWs into living cells using scalable processes has not yet been achieved. Here, NWs are inserted into living Chlamydomonas reinhardtii cells (Chlamy cells) via inkjet printing of the Chlamy cells, representing a low-cost and large-scale method for inserting NWs into living cells. Jetting conditions and printable bioink composed of living Chlamy cells are optimized to achieve stable jetting and precise ink deposition of bioink for indentation of NWs into Chlamy cells. Fluorescence confocal microscopy is used to verify the viability of Chlamy cells after inkjet printing. Simple mechanical considerations of the cell membrane and droplet kinetics are developed to control the jetting force to allow penetration of the NWs into cells. The results suggest that inkjet printing is an effective, controllable tool for stable insertion of NWs into cells with economic and scale-related advantages.

Prolonged and highly efficient intracellular extraction of photosynthetic electrons from single algal cells by optimized nanoelectrode insertion

Harvesting photosynthetic electrons (PEs) from plant or algal cells can be a highly efficient and environmentally friendly way of generating renewable energy. Recent work on nanoelectrode insertion into algal cells has demonstrated the possibility to directly extract PEs from living algal cells with high efficiencies. However, the instability of the inserted cells limits the practicality of this technology. Here, the impact of nanoelectrode insertion on intracellular extraction of PEs is characterized with the goal of stabilizing algal cells after nanoelectrode insertion. Using nanoelectrodes <500 nm in diameter, algal cells remained stable for over one week after insertion and continued to provide PEs through direct extraction by the inserted nanoelectrodes. After nanoelectrode insertion, a photosynthetic current density of 6 mA·cm−2, which is several fold higher than the current densities attained using approaches based on isolated thylakoid membranes or photosystem I complexes, was observed in the dark and during illumination at various light intensities.

Thylakoid-Deposited Micro-Pillar Electrodes for Enhanced Direct Extraction of Photosynthetic Electrons

Photosynthesis converts solar energy to electricity in a highly efficient manner. Since only water is needed as fuel for energy conversion, this highly efficient energy conversion process has been rigorously investigated. In particular, photosynthetic apparatus, such as photosystem II (PSII), photosystem I (PSI), or thylakoids, have been isolated from various plants to construct bio-hybrid anodes. Although PSII or PSI decorated anodes have shown potentials, there still remain challenges, such as poor stability of PSII-based systems or need for electron donors other than water molecules of PSI-based systems. Thylakoid membranes are relatively stable after isolation and they contain all the necessary photosynthetic apparatus including the PSII and PSI. To increase electrical connections between thylakoids and anodes, nanomaterials such as carbon nanotubes, nanowires, nanoparticles, or graphene have been employed. However, since they rely on the secondary electrical connections between thylakoids and anodes; it is desired to achieve larger direct contacts between them. Here, we aimed to develop micro-pillar (MP) array anodes to maximize direct contact with thylakoids. The thylakoid morphology was analyzed and the MP array was designed to maximize direct contact with thylakoids. The performance of MP anodes and a photosynthetic fuel cell based on MP electrodes was demonstrated and analyzed.

Scalable long-term extraction of photosynthetic electrons by simple sandwiching of nanoelectrode array with densely-packed algal cell film

Direct extraction of photosynthetic electrons from the whole photosynthetic cells such as plant cells or algal cells can be highly efficient and sustainable compared to other approaches based on isolated photosynthetic apparatus such as photosystems I, II, and thylakoid membranes. However, insertion of nanoelectrodes (NEs) into individual cells are time-consuming and unsuitable for scale-up processes. We propose simple and efficient insertion of massively-populated NEs into cell films in which algal cells are densely packed in a monolayer. After stacking the cell film over an NE array, gentle pressing of the stack allows a large number of NEs to be inserted into the cells in the cell film. The NE array was fabricated by metal-assisted chemical etching (MAC-etching) followed by additional steps of wet oxidation and oxide etching. The cell film was prepared by mixing highly concentrated algal cells with alginate hydrogel. Photosynthetic currents of up to 106 nA/cm2 was achieved without aid of mediators, and the photosynthetic function was maintained for 6 days after NE array insertion into algal cells.