Chia-Chang Tsai

Label-free detection of protein-protein interactions using a calmodulin-modified nanowire transistor

In this study, we describe a highly sensitive and reusable silicon nanowire field-effect transistor for the detection of protein-protein interactions. This reusable device was made possible by the reversible association of glutathione S-transferase-tagged calmodulin with a glutathione modified transistor. The calmodulin-modified transistor exhibited selective electrical responses to Ca2+ (≥1 μM) and purified cardiac troponin I (∼7 nM); the change in conductivity displayed a linear dependence on the concentration of troponin I in a range from 10 nM to 1 μM. These results are consistent with the previously reported concentration range in which the dissociation constant for the troponin I-calmodulin complex was determined. The minimum concentration of Ca2+ required to activate calmodulin was determined to be 1 μM. We have also successfully demonstrated that the N-type Ca2+ channels, expressed by cultured 293T cells, can be recognized specifically by the calmodulin-modified nanowire transistor. This sensitive nanowire transistor can serve as a high-throughput biosensor and can also substitute for immunoprecipitation methods used in the identification of interacting proteins.

High-Quality Graphene p−n Junctions via Resist-free Fabrication and Solution-Based Noncovalent Functionalization

An essential issue in graphene nanoelectronics is to engineer the carrier type and density and still preserve the unique band structure of graphene. We report the realization of high-quality graphene p-n junctions by noncovalent chemical functionalization. A generic scheme for the graphene p-n junction fabrication is established by combining the resist-free approach and spatially selective chemical modification process. The effectiveness of the chemical functionalization is systematically confirmed by surface topography and potential measurements, spatially resolved Raman spectroscopic imaging, and transport/magnetotransport measurements. The transport characteristics of graphene p-n junctions are presented with observations of high carrier mobilities, Fermi energy difference, and distinct quantum Hall plateaus. The chemical functionalization of graphene p-n junctions demonstrated in this study is believed to be a feasible scheme for modulating the doping level in graphene for future graphene-based nanoelectronics.

Exocytosis of a single bovine adrenal chromaffin cell: the electrical and morphological studies.

Exocytosis of a single bovine adrenal chromaffin cell, triggered by histamine stimulation, was investigated via the electric responses detected with single-walled carbon-nanotube field-effect transistors (SWCNT-FET) and the morphological changes acquired by atomic force microscopy (AFM). Secretion of chromogranin A (CgA), stored in the vesicles of a single chromaffin cell, can be monitored in situ by the antibody against CgA (CgA-antibody) functionalized on the SWCNT-FET devices. The SWCNT-FET can further discriminate the amount of released CgA with different levels of histamine stimulations. The AFM morphological studies on a chromaffin cell indicate that the depression structures on the cell surface, caused by the histamine-evoked exocytotic fusion pores, appeared much more frequently than those without histamine stimulation or with the pretreatment of mepyramine before histamine stimulation. The vesicle diameters are about 50 nm calculated from the obtained three-dimensional AFM images. In comparison, the fusion pores of chromaffin cells stimulated by high-K (+) buffer solution were also investigated to have a wider-ranging distribution of vesicle diameters of 60-260 nm. This work demonstrates that the combination of novel techniques, SWCNT-FET and AFM, can provide further insights into the fundamental properties of exocytosis in neuroendocrine cells.

Nanowire transistor-based ultrasensitive virus detection with reversible surface functionalization.

We have applied a reusable silicon nanowire field-effect transistor (SiNW-FET) as a biosensor to conduct ultrasensitive detection of H5N2 avian influenza virus (AIV) in very dilute solution. The reversible surface functionalization of SiNW-FET was made possible using a disulfide linker. In the surface functionalization, 3-mercaptopropyltrimethoxysilane (MPTMS) was first modified on the SiNW-FET (referred to as MPTMS/SiNW-FET), with subsequent dithiothreitol washing to reduce any possible disulfide bonding between the thiol groups of MPTMS. Subsequently, receptor molecules could be immobilized on the MPTMS/SiNW-FET by the formation of a disulfide bond. The success of the reversible surface functionalization was verified with fluorescence examination and electrical measurements. A surface topograph of the SiNW-FET biosensor modified with a monoclonal antibody against H5N2 virus (referred to as mAb(H5)/SiNW-FET) after detecting approximately 10(-17) M H5N2 AIVs was scanned by atomic force microscopy to demonstrate that the SiNW-FET is capable of detecting very few H5N2 AIV particles.

Dynasore inhibits rapid endocytosis in bovine chromaffin cells

Vesicle recycling is vital for maintaining membrane homeostasis and neurotransmitter release. Multiple pathways for retrieving vesicles fused to the plasma membrane have been reported in neuroendocrine cells. Dynasore, a dynamin GTPase inhibitor, has been shown to specifically inhibit endocytosis and vesicle recycling in nerve terminals. To characterize its effects in modulating vesicle recycling and repetitive exocytosis, changes in the whole cell membrane capacitance of bovine chromaffin cells were recorded in the perforated-patch configuration. Constitutive endocytosis was blocked by dynasore treatment, as shown by an increase in membrane capacitance. The membrane capacitance was increased during strong depolarizations and declined within 30 s to a value lower than the prestimulus level. The amplitude, but not the time constant, of the rapid exponential decay was significantly decreased by dynasore treatment. Although the maximal increase in capacitance induced by stimulation was significantly increased by dynasore treatment, the intercepts at time 0 of the curve fitted to the decay phase were all approximately 110% of the membrane capacitance before stimulation, regardless of the dynasore concentration used. Membrane depolarization caused clathrin aggregation and F-actin continuity disruption at the cell boundary, whereas dynasore treatment induced clathrin aggregation without affecting F-actin continuity. The number of invagination pits on the surface of the plasma membrane determined using atomic force microscopy was increased and the pore was wider in dynasore-treated cells. Our data indicate that dynamin-mediated endocytosis is the main pathway responsible for rapid compensatory endocytosis.

Chemical disorder-induced magnetism in FeSi2 nanoparticles

Iron disilicide in a bulk form is practically nonmagnetic. In contrast, nanoparticles of FeSi2 exhibit superparamagnetism with blocking temperatures ranging from 8K(15nm)to34K(55nm). Their relatively low saturation magnetization suggests that the magnetic behavior is associated with only a small fraction of Fe ions, which have a sufficient number of other Fe as nearest neighbors. The chemical disorder is presumably induced in the formation of nanoparticles. A spin glass-type anomaly below 10K observed in specific heat data gives a further evidence for the compositional heterogeneity.

Changes in Plasma Membrane Surface Potential of PC12 Cells as Measured by Kelvin Probe Force Microscopy

The plasma membrane of a cell not only works as a physical barrier but also mediates the signal relay between the extracellular milieu and the cell interior. Various stimulants may cause the redistribution of molecules, like lipids, proteins, and polysaccharides, on the plasma membrane and change the surface potential (Φs). In this study, the Φss of PC12 cell plasma membranes were measured by atomic force microscopy in Kelvin probe mode (KPFM). The skewness values of the Φss distribution histogram were found to be mostly negative, and the incorporation of negatively charged phosphatidylserine shifted the average skewness values to positive. After being treated with H2O2, dopamine, or Zn2+, phosphatidylserine was found to be translocated to the membrane outer leaflet and the averaged skewness values were changed to positive values. These results demonstrated that KPFM can be used to monitor cell physiology status in response to various stimulants with high spatial resolution.

Magnetic and superconducting properties of single crystals of Sr2HoRu1−xCuxO6 grown from high temperature solutions

Single crystals of Sr2HoRu1−xCuxO6 (with x=0–0.2), measuring 2–3mm across have been grown from PbO–PbF2 based solutions in the temperature range of 1250–1150°C. The crystals exhibit octahedral morphology and belong to the monoclinic space group P21∕n. While Sr2HoRuO6 is found to be antiferromagnetic with weak ferromagnetism below 30K, the solid solutions containing Cu exhibit a diamagnetic transition at 31K which increases in magnitude and temperature with increasing Cu. Through a correlation of magnetic and calorimetric properties, these crystals are concluded to be spin-glass superconductors.