Fenni Zhang

Impedance sensing and molecular modeling of an olfactory biosensor based on chemosensory proteins of honeybee.

By mimicking biological olfaction, biosensors have been used for the detection of important ligands in complex environments. An olfactory biosensor based on chemosensory proteins (CSPs) was designed by immobilizing honeybee CSPs (Ac-ASP3) on the interdigitated golden electrodes. Its responses to ligands of pheromones and floral odors were recorded by impedance spectroscopy. The relative decrease of charge transfer resistance of the biosensor is proportional to the logarithm of ligand concentration from 10(-7)M to 10(-3)M. To explore the molecular recognition processes of the biosensor, the tertiary structure of the protein was modeled and the protein-ligand interactions were investigated by the molecular docking. Our docking results verified the validity of experiments and showed that the specific ligands could form hydrogen bonds with some of the conserved residues, such as Cys 60 and Gln 64 of Ac-ASP3. Furthermore, combining the molecular modeling with impedance detection, the accuracy, specificity and predictability of the ligands binding to the protein could be improved. Thus, CSPs will provide a promising approach for chemical molecular sensing at low concentrations.

Bioelectronic tongue of taste buds on microelectrode array for salt sensing.

Taste has received great attention for its potential applications. In this work, we combine the biological tissue with micro-chips to establish a novel bioelectronic tongue system for salt taste detection. Before experiment, we established a computational model of action potential in salt taste receptor cell, simulating the responsive results to natural salt stimuli of NaCl solution with various concentrations. Then 36-channel microelectrode arrays (MEA) with the diameter of 30 μm were fabricated on the glass substrate, and taste epithelium was stripped from rat and fixed on MEA. When stimulated by the salt stimuli, electrophysiological activities of taste receptor cells in taste buds were measured through a multi-channel recording system. Both simulation and experiment results showed a dose-dependent increase in NaCl-induced potentials of taste receptor cells, which indicated good applications in salt measurements. The multi-channel analysis demonstrated that different groups of MEA channels were activated during stimulations, indicating non-overlapping populations of receptor cells in taste buds involved in salt taste perception. The study provides an effective and reliable biosensor platform to help recognize and distinguish salt taste components.

Extracellular recording of spatiotemporal patterning in response to odors in the olfactory epithelium by microelectrode arrays.

In olfactory biosensors, microelectronic sensor chips combined with biological olfactory cells are a promising platform for odor detection. In our investigation, olfactory epithelium stripped from rat was fixed on the surface of microelectrode arrays (MEAs). Electrophysiological activities of olfactory receptor neurons in intact epithelium were measured in the form of extracellular potentials. Based on multi-channel recording performance of MEA and structural and functional integrality of native olfactory epithelium, the spatiotemporal analysis was carried out to study the extracellular activity pattern of neurons in the tissue. The variation of spatiotemporal patterns corresponding to different odors displayed the signals firing image characteristic intuitionally. It is an effective method in the form of patterns for monitoring the state of tissue both in time and space domain, promoting the platform for olfactory sensing mechanism research.

Extracellular potentials recording in intact taste epithelium by microelectrode array for a taste sensor.

Taste receptor cells in taste buds can generate action potentials in response to taste stimuli. The spatiotemporal patterns of the potentials have great value in both biomedical and engineering researches. In the present study, by fixing the biological epithelium onto the surface of microelectrode arrays (MEA), we established a novel taste sensor to record action potentials from the taste receptor cells of rat in response to taste stimuli. By this multi-channel recording method, we examined the electrophysiological activities of taste receptor cells in taste buds to stimuli representing the basic taste qualities (sour, salt, bitter, sweet and umami). The recorded action potentials corresponding to five tastes displayed different spatiotemporal patterns. The multi-channel results demonstrated that taste buds released the spontaneous signals simultaneously and displayed different responses to different tastes stimulations. The temporal characteristics were derived by time-domain and frequency-domain analysis, and the signals fired in different stimuli could be distinguished into different clusters by principal component analysis (PCA). The study provides an effective and reliable platform to recognize and distinguish basic taste qualities.

Biosensor analysis of natural and artificial sweeteners in intact taste epithelium

Sweeteners are commonly used as food additives in our daily life, which, however, have been causing a number of undesirable diseases since the last century. Therefore, the detection and quantification of sweeteners are of great value for food safety. In this study, we used a taste biosensor to measure and analyze different sweeteners, both natural and artificial sweeteners included. Electrophysiological activities from taste epithelium were detected by the multi-channel biosensors and analyzed with spatiotemporal methods. The longtime signal result showed different temporal-frequency properties with stimulations of individual sweeteners such as glucose, sucrose, saccharin, and cyclamate, while the multi-channel results in our study revealed the spatial expression of taste epithelium to sweet stimuli. Furthermore, in the analysis of sweetener with different concentrations, the result showed obvious dose-dependent increases in signal responses of the taste epithelium, which indicated promising applications in sweetness evaluation. Besides, the mixture experiment of two natural sweeteners with a similar functional unit (glucose and sucrose) presented two signal patterns, which turned out to be similar with responses of each individual stimulus involved. The biosensor analysis of common sweeteners provided new approaches for both natural and artificial sweeteners evaluation.

Olfactory epithelium biosensor: odor discrimination of receptor neurons from a bio-hybrid sensing system

Bio-hybrid systems provide an opportunity for integrating a living bio-active unit and a proper biosensing system, to employ the unique properties of the bio-active unit. The biological olfactory system can sense and identify thousands of trace odors. The purpose of this study is to combine olfactory epithelium with microelectrode array (MEA) to establish an olfactory epithelium-MEA hybrid system to record the odor-induced electrophysiological activities of the tissue. In our experiments, extracellular potential of olfactory receptor neurons in intact epithelium were measured in the presence of ethyl ether, acetic acid, butanedione, and acetone, respectively. After the odor-induced response signals were analyzed in the time and frequency domain, the temporal characteristics of response signals were extracted. We found that olfactory epithelium-MEA hybrid system can reflect the in vitro odor information of different signal characteristics and firing modes in vitro. The bio-hybrid sensing system can represent a useful instrument to sense and detect the odorant molecules with well recognizing patterns. With the development of sensor technology, bio-hybrid systems will represent emerging and promising platforms for wide applications, ranging from health care to environmental monitoring.

Umami evaluation in taste epithelium on microelectrode array by extracellular electrophysiological recording

Umami is one of the basic tastes along with sweet, bitter, sour and salty. It is often elicited by amino acids and can provide a palatable flavor for food. With taste epithelium as the sensing element, microelectrodes can be used to evaluate umami taste by biological responses of the tissue. The electrophysiological activities to umami stimuli are measured with a 60-channel microelectrode array (MEA). Local field potential (LFP) recorded by a MEA system showed different temporal characteristics respectively with l-glutamic acid (l-Glu), l-aspartic acid (l-Asp), l-monosodium glutamate (l-MSG) and l-monosodium aspartate (l-MSA), while remarkable differences were observed between amino acids and their sodium salts. We also found that a dose-dependent behavior in the increasing concentrations of umami stimulations and a synergistic enhancement between amino acids and purine nucleotides can be detected. The investigation of this evaluation for umami represents a promising approach for distinguishing and evaluating umami tastants.

Neurosecretory cell-based biosensor: Monitoring secretion of adrenal chromaffin cells by local extracellular acidification using light-addressable potentiometric sensor

Vesicular exocytosis plays an important role in many physiological processes. The dense-core vesicles release of chromaffin cells is a suitable model for the presynaptic process in neurosecretory cells. In this study, light-addressable potentiometric sensor (LAPS) was introduced as a label-free recording method for vesicle release by the local extracellular acidification. The chromaffin cells are directly cultured on the sensor surface. After cells and LAPS hybrid system is established, the events of vesicular exocytosis are recorded. Protons stored in the vesicles and co-released with transmitters, induced a brief acidic shifts in the cell-sensor cleft. Under the stimulation of the KCl and acetylcholine (Ach), the signals presented the different amplitude and exocytosis rate, and reflected the specific features of the exocytosis. The result indicates that neurosecretory cell-based biosensor will provide a useful platform for neurosecretion mechanism research by monitoring the exocytotic activities with extracellular acidification sensing.

Microelectrode recording of tissue neural oscillations for a bionic olfactory biosensor

In olfactory research, neural oscillations exhibit excellent temporal regularity, which are functional and necessary at the physiological and cognitive levels. In this paper, we employed a bionic tissue biosensor which treats intact epithelium as sensing element to record the olfactory oscillations extracellularly. After being stimulated by odorant of butanedione, the olfactory receptor neurons generated different kinds of oscillations, which can be described as pulse firing oscillation, transient firing oscillation, superposed firing oscillation, and sustained firing oscillation, according to their temporal appearances respectively. With a time-frequency analysis of sonogram, the oscillations also demonstrated different frequency properties, such as δ, θ, α, β and γ oscillations. The results suggest that the bionic biosensor cooperated with sonogram analysis can well improve the investigation of olfactory oscillations, and provide a novel model for artificial olfaction sensor design.

Odors Discrimination by Olfactory Epithelium Biosensor

Humans are exploring the bionic biological olfaction to sense the various trace components of gas or liquid in many fields. For achieving the goal, we endeavor to establish a bioelectronic nose system for odor detection by combining intact bioactive function units with sensors. The bioelectronic nose is based on the olfactory epithelium of rat and microelectrode array (MEA). The olfactory epithelium biosensor generates extracellular potentials in presence of odor, and presents obvious specificity under different odors condition. The odor response signals can be distinguished with each other effectively by signal sorting. On basis of bioactive MEA hybrid system and the improved signal processing analysis, the bioelectronic nose will realize odor discrimination by the specific feature of signals response to various odors.