Recent Advances in Electrochemiluminescence Visual Biosensing and Bioimaging.

Electrochemiluminescence (ECL) is one of the most powerful techniques that meet the needs of analysis and detection in a variety of scenarios, because of its highly analytical sensitivity and excellent spatiotemporal controllability. ECL combined with microscopy (ECLM) offers a promising approach for quantifying and mapping a wide range of analytes. To date, ECLM has been widely used to image biological entities and processes, such as cells, subcellular structures, proteins and membrane transport properties. In this review, we first introduced the mechanisms of several classic ECL systems, then highlighted the progress of visual biosensing and bioimaging by ECLM in the last decade. Finally, the characteristics of ECLM were summarized, as well as some of the current challenges. The future research interests and potential directions for the application of ECLM were also outlooked.

T Cell Antigen Recognition and Discrimination by Electrochemiluminescence Imaging.

Adoptive T lymphocyte (T cell) transfer and tumour-specific peptide vaccines are innovative cancer therapies. An accurate assessment of the specific reactivity of T cell receptors (TCRs) to tumour antigens is required because of the high heterogeneity of tumour cells and the immunosuppressive tumour microenvironment. In this study, we report a label-free electrochemiluminescence (ECL) imaging approach for recognising and discriminating between TCRs and tumour-specific antigens by imaging the immune synapses of T cells. Various T cell stimuli, including agonistic antibodies, auxiliary molecules, and tumour-specific antigens, were modified on the electrode's surface to allow for their interaction with T cells bearing different TCRs. The formation of immune synapses activated by specific stimuli produced a negative (shadow) ECL image, from which T cell antigen recognition and discrimination were evaluated by analysing the spreading area and the recognition intensity of T cells. This approach provides an easy way to assess TCR-antigen specificity and screen both of them for immunotherapies.

Recent advances in label-free imaging of cell-matrix adhesions.

Cell-matrix adhesions play an essential role in mediating and regulating many biological processes. The adhesion receptors, typically transmembrane integrins, provide dynamic correlations between intracellular environments and extracellular matrixes (ECMs) by bi-directional signaling. In-depth investigations of cell-matrix adhesion and integrin-mediated cell adhesive force are of great significance in biology and medicine. The emergence of advanced imaging techniques and principles has facilitated the understanding of the molecular composition and structure dynamics of cell-matrix adhesions, especially the label-free imaging methods that can be used to study living cell dynamics without immunofluorescence staining. This highlight article aims to give an overview of recent developments in imaging cell-matrix adhesions in a label-free manner. Electrochemiluminescence microscopy (ECLM) and surface plasmon resonance microscopy (SPRM) are briefly introduced and their applications in imaging analysis of cell-matrix adhesions are summarized. Then we highlight the advances in mapping cell-matrix adhesion force based on molecular tension probes and fluorescence microscopy (collectively termed as MTFM). The biomaterials including polyethylene glycol (PEG), peptides and DNA for constructing tension probes in MTFM are summarized. Finally, the outlook and perspectives on the further developments of cell-matrix adhesion imaging are presented.

Surface-Sensitive Imaging Analysis of Cell-Microenvironment Interactions by Electrochemiluminescence Microscopy.

A complex and heterogeneous cell microenvironment offers not only structural support for cells but also myriad biochemical and biophysical cues. These outside-in signals transmit into cells primarily through integrins, which are the important components of cell-matrix adhesions to direct and maintain cell behaviors and fate. In this work, we report a surface-sensitive imaging methodology for evaluating the difference in cell-matrix adhesions at the single cell level to dissect the impact of the chemical microenvironment on cell behaviors. Cells were cultured on silica nanochannel membrane (SNM) modified indium tin oxide (ITO) electrodes (SNM/ITO) with different terminal surfaces and imaged by electrochemiluminescence microscopy (ECLM). The results show that the surface tethered with Arg-Gly-Asp (RGD) groups can mediate robust cell-microenvironment interaction and those coated with silanol and (3-aminopropyl)triethoxysilane (APTES) groups transmit an intermediate adhesion, while oligo(ethylene glycol) (OEG) coated surface conveys the weakest cell-matrix adhesion. Specific recognition of integrins to different surfaces was further explored in conjunction with selective immunoblocking of different subunits. α6, α5, and α1 integrin subunits were found to recognize SNM, RGD/OEG, and APTES surfaces, respectively. The work provides not only insights into cell-microenvironment interaction but also guideline in the design and development of functional and biomimetic surface materials.

Spatially Selective Imaging of Cell-Matrix and Cell-Cell Junctions by Electrochemiluminescence.

Cell junctions are protein structures located at specific cell membrane domains that determine key processes in multicellular development. Here we report spatially selective imaging of cell junctions by electrochemiluminescence (ECL) microscopy. By regulating the concentrations of luminophore and/or co-reactant, the thickness of ECL layer can be controlled to match with the spatial location of different cell junctions. At a low concentration of luminophore, ECL generation is confined to the electrode surface, thus revealing only cell-matrix adhesions at the bottom of cells. While at a high concentration of luminophore, the ECL layer can be remarkably extended by decreasing the co-reactant concentration, thus allowing the sequential imaging of cell-matrix and cell-cell junctions at the bottom and near the apical surface of cells, respectively. This strategy not only provides new insights into the ECL mechanisms but also promises wide applications of ECL microscopy in bioimaging.

Quantum transport in Chern insulators on Möbius strips.

Chern insulators (CIs) or quantum anomalous Hall (QAH) states have drawn more attention, with emergence of quantized Hall conductance but in absence of Landau levels. Here, we study the Haldane-type CI/QAH states on Möbius strips and focus on the quantum transport properties. The Möbius strips can be constructed from the twisted honeycomb-lattice strips with domain walls. Topological properties of the Möbius CI/QAH states can be identified by the local density of states, the real-space Chern numbers and the transport properties. Nearly perfect quantized conductance plateaus are observed in the numerical quantum-transport simulations. In addition, we found that adding magnetic flux in domain walls can induce alternating edge states. At last, we discussed in details the different lead connection schemes, and found that we can use multiple domain walls to obtain various higher quantized conductance plateaus.