Member (Student), Cardiovascular Institute
Honors & Awards
Dean Award (Top 10% Outstanding students), College of Science, National Taiwan University (2017)
45th International Chemistry Olympiad Gold Prize, Russia (2013)
Education & Certifications
B.S., National Taiwan University, Chemistry (2017)
Expansion microscopy for imaging the cell-material interface.
2023; 122 (3S1): 133a
View details for DOI 10.1016/j.bpj.2022.11.883
View details for PubMedID 36782597
Modulation of nuclear membrane repair machinery by nano-needle arrays.
2023; 122 (3S1): 552a
View details for DOI 10.1016/j.bpj.2022.11.2922
View details for PubMedID 36784865
A versatile nanoelectrode platform for electrical recording of diverse cell types.
2023; 122 (3S1): 431a
View details for DOI 10.1016/j.bpj.2022.11.2333
View details for PubMedID 36784209
Engineering cell morphology using maskless 2D protein micropatterning on 3D nanostructures.
2023; 122 (3S1): 553a
View details for DOI 10.1016/j.bpj.2022.11.2925
View details for PubMedID 36784871
Cardiotoxicity drug screening based on whole-panel intracellular recording.
Biosensors & bioelectronics
2022; 216: 114617
Unintended binding of small-molecule drugs to ion channels affects electrophysiological properties of cardiomyocytes and potentially leads to arrhythmia and heart failure. The waveforms of intracellular action potentials reflect the coordinated activities of cardiac ion channels and serve as a reliable means for assessing drug toxicity, but the implementation is limited by the low throughput of patch clamp for intracellular recording measurements. In the last decade, several new technologies are being developed to address this challenge. We recently developed the nanocrown electrode array (NcEA) technology that allows robust, parallel, and long-duration recording of intracellular action potentials (iAPs). Here, we demonstrate that NcEAs allow comparison of iAP waveforms before and after drug treatment from the same cell. This self-referencing comparison not only shows distinct drug effects of sodium, potassium, and calcium blockers, but also reveals subtle differences among three subclasses of sodium channel blockers with sub-millisecond accuracy. Furthermore, self-referencing comparison unveils heterogeneous drug responses among different cells. In our study, whole-panel simultaneous intracellular recording can be reliably achieved with 94% success rate. The average duration of intracellular recording is 30min and some last longer than 2h. With its high reliability, long recording duration, and easy-to-use nature, NcEA would be useful for iAP-based preclinical drug screening.
View details for DOI 10.1016/j.bios.2022.114617
View details for PubMedID 36027802
Expansion Microscopy for Imaging the Cell-Material Interface.
Surface topography on the scale of tens of nanometers to several micrometers substantially affects cell adhesion, migration, and differentiation. Recent studies using electron microscopy and super-resolution microscopy provide insight into how cells interact with surface nanotopography; however, the complex sample preparation and expensive imaging equipment required for these methods makes them not easily accessible. Expansion microscopy (ExM) is an affordable approach to image beyond the diffraction limit, but ExM cannot be readily applied to image the cell-material interface as most materials do not expand. Here, we develop a protocol that allows the use of ExM to resolve the cell-material interface with high resolution. We apply the technique to image the interface between U2OS cells and nanostructured substrates as well as the interface between primary osteoblasts with titanium dental implants. The high spatial resolution enabled by ExM reveals that although AP2 and F-actin both accumulate at curved membranes induced by vertical nanostructures, they are spatially segregated. Using ExM, we also reliably image how osteoblasts interact with roughened titanium implant surfaces below the diffraction limit; this is of great interest to understand osseointegration of the implants but has up to now been a significant technical challenge due to the irregular shape, the large volume, and the opacity of the titanium implants that have rendered them incompatible with other super-resolution techniques. We believe that our protocol will enable the use of ExM as a powerful tool for cell-material interface studies.
View details for DOI 10.1021/acsnano.1c11015
View details for PubMedID 35533401
Nanocrown electrodes for parallel and robust intracellular recording of cardiomyocytes.
2022; 13 (1): 2253
Drug-induced cardiotoxicity arises primarily when a compound alters the electrophysiological properties of cardiomyocytes. Features of intracellular action potentials (iAPs) are powerful biomarkers that predict proarrhythmic risks. In the last decade, a number of vertical nanoelectrodes have been demonstrated to achieve parallel and minimally-invasive iAP recordings. However, the large variability in success rate and signal strength have hindered nanoelectrodes from being broadly adopted for proarrhythmia drug assessment. In this work, we develop vertically-aligned nanocrown electrodes that are mechanically robust and achieve>99% success rates in obtaining intracellular access through electroporation. We validate the accuracy of nanocrown electrode recordings by simultaneous patch clamp recording from the same cell. Finally, we demonstrate that nanocrown electrodes enable prolonged iAP recording for continual monitoring of the same cells upon the sequential addition of four incremental drug doses. Our technology development provides an advancement towards establishing an iAP screening assay for preclinical evaluation of drug-induced arrhythmogenicity.
View details for DOI 10.1038/s41467-022-29726-2
View details for PubMedID 35474069
Exploring cell-surface nanopillar interactions with 3D superresolution microscopy
CELL PRESS. 2022: 278A
View details for Web of Science ID 000759523001617
Intracellular recording of cardiac action potentials via membrane electroporation
CELL PRESS. 2022: 304A
View details for Web of Science ID 000759523002022
Mesh electrode arrays for integration with electrogenic organoids
CELL PRESS. 2022: 16
View details for Web of Science ID 000759523000078
Membrane curvature regulates the spatial distribution of bulky glycoproteins
View details for DOI 10.1038/s41467-022-30610-2
Nanoscale Surface Topography Reduces Focal Adhesions and Cell Stiffness by Enhancing Integrin Endocytosis.
Both substrate stiffness and surface topography regulate cell behavior through mechanotransduction signaling pathways. Such intertwined effects suggest that engineered surface topographies might substitute or cancel the effects of substrate stiffness in biomedical applications. However, the mechanisms by which cells recognize topographical features are not fully understood. Here we demonstrate that the presence of nanotopography drastically alters cell behavior such that neurons and stem cells cultured on rigid glass substrates behave as if they were on soft hydrogels. With atomic force microscopy, we show that rigid nanotopography resembles the effects of soft hydrogels in reducing cell stiffness and membrane tension. Further, we reveal that nanotopography reduces focal adhesions and cell stiffness by enhancing the endocytosis and the subsequent removal of integrin receptors. This mechanistic understanding will support the rational design of nanotopography that directs cells on rigid materials to behave as if they were on soft ones.
View details for DOI 10.1021/acs.nanolett.1c01934
View details for PubMedID 34346220
Exploring Cell Surface-Nanopillar Interactions with 3D Super-Resolution Microscopy.
Plasma membrane topography has been shown to strongly influence the behavior of many cellular processes such as clathrin-mediated endocytosis, actin rearrangements, and others. Recent studies have used three-dimensional (3D) nanostructures such as nanopillars to imprint well-defined membrane curvatures (the "nano-bio interface"). In these studies, proteins and their interactions were probed by two-dimensional fluorescence microscopy. However, the low resolution and limited axial detail of such methods are not optimal to determine the relative spatial position and distribution of proteins along a 100 nm-diameter object, which is below the optical diffraction limit. Here, we introduce a general method to explore the nanoscale distribution of proteins at the nano-bio interface with 10-20 nm precision using 3D single-molecule super-resolution (SR) localization microscopy. This is achieved by combining a silicone-oil immersion objective and 3D double-helix point spread function microscopy. We carefully adjust the objective to minimize spherical aberrations between quartz nanopillars and the cell. To validate the 3D SR method, we imaged the 3D shape of surface-labeled nanopillars and compared the results with electron microscopy measurements. Turning to transmembrane-anchored labels in cells, the high quality 3D SR reconstructions reveal the membrane tightly wrapping around the nanopillars. Interestingly, the cytoplasmic protein AP-2 involved in clathrin-mediated endocytosis accumulates along the nanopillar above a specific threshold of 1/R (the reciprocal of the radius) membrane curvature. Finally, we observe that AP-2 and actin preferentially accumulate at positive Gaussian curvature near the pillar caps. Our results establish a general method to investigate the nanoscale distribution of proteins at the nano-bio interface using 3D SR microscopy.
View details for DOI 10.1021/acsnano.1c05313
View details for PubMedID 34582687
Quantitative Nano-Platforms for Interrogation of Curvature Sensitive Proteins
CELL PRESS. 2020: 249A–250A
View details for Web of Science ID 000513023201503