INSULIN secretion was traditionally measured with biochemical and immunological methods such as enzyme linked immunosorbant assay and radioimmunoassay. However, these methods can only tell the amount of insulin secret...INSULIN secretion was traditionally measured with biochemical and immunological methods such as enzyme linked immunosorbant assay and radioimmunoassay. However, these methods can only tell the amount of insulin secreted; they give no information about the secretion process or mechanism of exocytosis. In recent years, an imaging technique known as total internal reflection fluorescence （TIRF） microscopy has been employed to study insulin secretion.展开更多
A spectrometric setup to perform total internal reflection fluorescence (TIRF) and synchronous TIRF measurements at solid/liquid interfaces is presented. The combination of TIRF and synchronous fluorescence was propo...A spectrometric setup to perform total internal reflection fluorescence (TIRF) and synchronous TIRF measurements at solid/liquid interfaces is presented. The combination of TIRF and synchronous fluorescence was proposed to analyze simultaneously different components at interfaces. The TIRF excitation, emission and synchronous spectra of a watersoluble porphyrin were obtained from water/glass interface using this setup without the existence of a surfactant.展开更多
Total internal reflection fluorescence spectroscopy (TIRF) and synchronous scanning technique were combined to study the adsorption behavior of the meso-tetrakis (4-sulfonatophenyl) porphyrin (TPPS) at the glass-water...Total internal reflection fluorescence spectroscopy (TIRF) and synchronous scanning technique were combined to study the adsorption behavior of the meso-tetrakis (4-sulfonatophenyl) porphyrin (TPPS) at the glass-water interface without any surfactant. The pH dependence of synchronous fluorescence signal at the interface was analyzed. Both unprotonated (TPPS4-) and diprotonated (H2TPPS2-) forms of TPPS were observed at the interface. But the interface favored the adsorption of. The apparent estimated pKa2 value shifted from 5.00 in the bulk solution to 2.7 at the interface. STIRF provides a good technique to study multi-component systems at the interface.展开更多
Fluorescence microscopy has evolved from a purely biological tool to a powerful chemical instrument for imaging and kinetics research into nanocatalysis.And the demand for high signal-to-noise ratio and temporal–spat...Fluorescence microscopy has evolved from a purely biological tool to a powerful chemical instrument for imaging and kinetics research into nanocatalysis.And the demand for high signal-to-noise ratio and temporal–spatial resolution detection has encouraged rapid growth in total internal reflection fluorescence microscopy(TIRFM).By producing an evanescent wave on the glass–water interface,excitation can be limited to a thin plane to ensure the measured accuracy of kinetics and image contrast of TIRFM.Thus,this unique physical principle of TIRFM makes it suitable for chemical research.This review outlines applications of TIRFM in the field of chemistry,including imaging and kinetics research.Hence,this review could provide guidance for beginners employing TIRFM to solve current challenges creatively in chemistry.展开更多
Adhesion molecule CD146 (100-130kDa) belongs to the immunoglobulin super family and it is originally identified as a biomarker for melanoma. Recently, CD146 is found as
Fluorescence imaging of single molecules is becoming a powerful tool to examine biological processes at the molecular level.Using total internal reflection fluorescence microscopy (TIRFM),it has been possible to study...Fluorescence imaging of single molecules is becoming a powerful tool to examine biological processes at the molecular level.Using total internal reflection fluorescence microscopy (TIRFM),it has been possible to study the dynamic behavior of single molecules on living cell membranes.Herein,we briefly review the application of TIRFM-based single-molecule imaging in studies of membrane receptors involved in signal transduction.Furthermore,we discuss several examples of our own research on growth factor receptors,including TGF-β receptors,HER2,and EGFR,and speculate possible applications of this technique to investigate other cellular events occurring on or near the plasma-membrane.展开更多
文摘INSULIN secretion was traditionally measured with biochemical and immunological methods such as enzyme linked immunosorbant assay and radioimmunoassay. However, these methods can only tell the amount of insulin secreted; they give no information about the secretion process or mechanism of exocytosis. In recent years, an imaging technique known as total internal reflection fluorescence （TIRF） microscopy has been employed to study insulin secretion.
文摘A spectrometric setup to perform total internal reflection fluorescence (TIRF) and synchronous TIRF measurements at solid/liquid interfaces is presented. The combination of TIRF and synchronous fluorescence was proposed to analyze simultaneously different components at interfaces. The TIRF excitation, emission and synchronous spectra of a watersoluble porphyrin were obtained from water/glass interface using this setup without the existence of a surfactant.
文摘Total internal reflection fluorescence spectroscopy (TIRF) and synchronous scanning technique were combined to study the adsorption behavior of the meso-tetrakis (4-sulfonatophenyl) porphyrin (TPPS) at the glass-water interface without any surfactant. The pH dependence of synchronous fluorescence signal at the interface was analyzed. Both unprotonated (TPPS4-) and diprotonated (H2TPPS2-) forms of TPPS were observed at the interface. But the interface favored the adsorption of. The apparent estimated pKa2 value shifted from 5.00 in the bulk solution to 2.7 at the interface. STIRF provides a good technique to study multi-component systems at the interface.
基金This work was supported by the National Science Foundation of China(21925205,22072145,22102172,21721003)。
文摘Fluorescence microscopy has evolved from a purely biological tool to a powerful chemical instrument for imaging and kinetics research into nanocatalysis.And the demand for high signal-to-noise ratio and temporal–spatial resolution detection has encouraged rapid growth in total internal reflection fluorescence microscopy(TIRFM).By producing an evanescent wave on the glass–water interface,excitation can be limited to a thin plane to ensure the measured accuracy of kinetics and image contrast of TIRFM.Thus,this unique physical principle of TIRFM makes it suitable for chemical research.This review outlines applications of TIRFM in the field of chemistry,including imaging and kinetics research.Hence,this review could provide guidance for beginners employing TIRFM to solve current challenges creatively in chemistry.
文摘Adhesion molecule CD146 (100-130kDa) belongs to the immunoglobulin super family and it is originally identified as a biomarker for melanoma. Recently, CD146 is found as
基金supported by the National Basic Research Program of China(2007CB935601)the National Natural Science Foundation of China(90713024,20821003)the Chinese Academy of Sciences
文摘Fluorescence imaging of single molecules is becoming a powerful tool to examine biological processes at the molecular level.Using total internal reflection fluorescence microscopy (TIRFM),it has been possible to study the dynamic behavior of single molecules on living cell membranes.Herein,we briefly review the application of TIRFM-based single-molecule imaging in studies of membrane receptors involved in signal transduction.Furthermore,we discuss several examples of our own research on growth factor receptors,including TGF-β receptors,HER2,and EGFR,and speculate possible applications of this technique to investigate other cellular events occurring on or near the plasma-membrane.
