Pingang He

A scanning electrochemical microscope (SECM) combined with an Au–Cu dual-electrode tip was used to in situ monitor the effect of Cu ²⁺ on the membrane permeability of a single living cell.

The imaging resolution of scanning electrochemical microscopy (SECM) depends strongly on the tip electrode size and the tip–substrate distance. Herein, etched glass encapsulation was applied to fabricate a gold disk electrode, and the size of the tip electrode was accurately determined from the steady-state limiting current. Referring to the theoretical research carried out by our predecessors, the formula for the imaging resolution was derived, followed by the imaging of gold spots and cells with the prepared microelectrodes of different sizes and with different tip–substrate distances. A depth scan was performed to generate 2D current maps of the gold spot relative to the position of the microelectrode in the x–z plane. Probe approach curves and horizontal sweeps were obtained from one depth scan image by simply extracting vertical and horizontal cross-sectional lines, and further characterized by comparison with simulated curves through modeling of the experimental system. The experimental results were basically consistent with the theory, revealing that the highest imaging resolution can be obtained with the smallest tip electrode when d/a = 1, and when the size of the tip electrode is fixed the smallest tip–substrate distance can give the highest imaging resolution.

A programmable multi-target-response electrochemical imaging technique was presented using scanning electrochemical microscopy (SECM) combining with self-designed waveform. The potential waveform applied to the tip decreased the charging current caused by potential switch, enhancing the signal-to-noise ratio. This programmable SECM (P-SECM) method was used to scan a metal strip for verifying its feasibility in feedback mode. Since it could achieve simultaneous multi-target imaging during one single imaging process, PC12 cells status was imaged and identified through three different molecules (FcMeOH, Ru(NH3)63+ and oxygen). The FcMeOH image eliminated the error from cell height and the Ru(NH3)63+ image verified the change of membrane permeability. Moreover, the oxygen image demonstrated the bioactivity of the cell via its intensity of respiration. Combining information from these three molecules, cell status could be determined accurately and also the error caused by time consumption with multiple scans in traditional SECM was eliminated.

Herein we propose a multiple signal amplification strategy designed for ultrasensitive electrochemical detection of glycoproteins. This approach introduces a new type of boronate-affinity sandwich assay (BASA), which was fabricated by using gold nanoparticles combined with reduced graphene oxide (AuNPs-GO) to modify sensing surface for accelerating electron transfer, the composite of molecularly imprinted polymer (MIP) including 4-vinylphenylboronic acid (VPBA) for specific capturing glycoproteins, and SiO₂ nanoparticles carried gold nanoparticles (SiO₂@Au) labeled with 6-ferrocenylhexanethiol (FcHT) and 4-mercaptophenylboronic acid (MPBA) (SiO₂@Au/FcHT/MPBA) as tracing tag for binding glycoprotein and generating electrochemical signal. As a sandwich-type sensing, the SiO₂@Au/FcHT/MPBA was captured by glycoprotein on the surface of imprinting film for further electrochemical detection in 0.1 M PBS (pH 7.4). Using horseradish peroxidase (HRP) as a model glycoprotein, the proposed approach exhibited a wide linear range from 1 pg/mL to 100 ng/mL, with a low detection limit of 0.57 pg/mL. To the best of our knowledge, this is first report of a multiple signal amplification approach based on boronate-affinity molecularly imprinted polymer and SiO₂@Au/FcHT/MPBA, exhibiting greatly enhanced sensitivity for glycoprotein detection. Furthermore, the newly constructed BASA based glycoprotein sensor demonstrated HRP detection in real sample, such as human serum, suggesting its promising prospects in clinical diagnostics.

Cell alignment plays a crucial role in the repair and regeneration of tissues, which is caused by the "contact guidance" of micro/nano structures. In this paper, the nano-grooves with 200 nm in width, 400 nm in period and 75 nm in depth were fabricated on gold substrate with the technique of nanoimprint to simulate the extracellular matrix (ECM). Electric cell-substrate impedance sensing (ECIS) was employed firstly to real-time monitor cell alignment of human foreskin fibroblasts (HFF) and human immortal keratinocyte cells (HaCaT) on nano-grooves, which are two important functional cell types in skin wound-healing. The cell images displayed that the nano-grooves could induce the alignment of HFF cells, in which, the cell arrangement along the direction of nano-grooves occurred prior to the cell elongation. While the nano-grooves couldn't influence the morphology of HaCaT cells, and their adhesion and spread were delayed. In the ECIS monitoring, HFF and HaCaT cells both presented increased normalized impedance (NI) values at their respective characteristic frequencies of 977 and 1465 Hz on nano-grooves and flat electrodes with the prolongation of culture time during 24 h and the increasing trends of NI values were also similar: in the first 6 h, NI values increased faster, and then, the increasing rates declined obviously. HFF cells on nano-grooves generated more intense impedance signals with a larger distinction of increasing rate than those on flat electrodes, indicating that the nano-grooves could promote the adhesion and spread of HFF cells and the directional arrangement had a larger impact on the variation of NI values than cell elongation. While HFF cells adhered and spread in random directions, leading to the reduced difference in increasing rate of NI values. The NI values of HFF cells was further correlated with cell alignment, showing the enhanced impedance responses with the rising percentage of aligned cells. More importantly, there was a good linear correlation between NI values and the percentage of cells arranging along the direction of nano-grooves. In contrast, HaCaT cells had smaller NI values on the nano-grooves with the similar increasing rates, compared to the flat electrodes, revealing that the nano-grooves were less suitable for the adhesion and spread of HaCaT cells with almost no change of cell morphology, and the cell adhesion could cause more obvious variation of NI values than cell spread. Our research would provide a support for the development of complex cell sensors based on ECIS and its application in clinical research field.

A new series of metallocyclodextrins with increased fluorescence intensity upon binding with ssDNAs/aptamers has been demonstrated to sensitively detect lysozyme.

Plasmonic nanoparticles have been widely applied in cell imaging, disease diagnosis, and photothermal therapy owing to their unique scattering and absorption spectra based on localized surface plasmon resonance (LSPR) property. Recently, it is still a big challenge to study the detailed scattering properties of single plasmonic nanoparticles in living cells and tissues, which have dynamic and complicated environment. The conventional approach for measuring the scattering light is based on a spectrograph coupled to dark-field microscopy (DFM), which is time-consuming and limited by the small sample capacity. Alternatively, RGB-based method is promising in high-throughput analysis of single plasmonic nanoparticles in dark-field images, but the limitation in recognition of nanoparticles hinders its application for intracellular analysis. In this paper, we developed an automatic and robust method for recognizing the plasmonic nanoparticles in dark-field image for RGB-based analysis. The method involves a bias-modified fuzzy C-means algorithm, through which biased illumination in the image could be eliminated. Thus, nearly all of the gold nanoparticles in the recorded image were recognized both on glass slide and in living cells. As confirmed, the distribution of peak wavelength obtained by our method is well agreed to the result measured by conventional method. Furthermore, we demonstrated that our method is profound in cell imaging studies, where its advantages in fast and high-throughput analysis of the plasmonic nanoparticles could be applied to confirm the presence and location of important biological molecules and provide efficiency information for cancer drug selection.