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A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions
Published on: November 23, 2015
In-situ and amplification-free imaging of hERG ion channels at single-cell level using a unique
Wen-Shu Zhang1, Xiao-Peng Liu1, Shuai Yue2
1Research Center for Analytical Sciences, Northeastern University, Shenyang, 110819, PR China.
Insights
Researchers developed novel gold-silver core-shell nanoparticles for surface-enhanced Raman scattering (SERS) to study the human ether-a-go-go related gene (hERG) potassium ion channel. This SERS approach enables in-situ imaging of ion channel distribution on cell surfaces.
Area of Science:
- Biophysics
- Nanotechnology
- Molecular Biology
Background:
- Ion channel research is crucial for drug development and disease diagnosis.
- Current methods often lack in vivo cell microenvironment relevance, limiting studies on cell structure-ion channel function links.
- Investigating ion channels in their native cellular context is essential for understanding their roles.
Purpose of the Study:
- To develop a novel surface-enhanced Raman scattering (SERS) nanoprobe for investigating ion channels.
- To study the human ether-a-go-go related gene (hERG) potassium ion channel in cell membranes.
- To establish an amplification-free method for in-situ imaging of ion channel distribution at the single-cell level.
Main Methods:
- Synthesis of gold core-4-mercaptobenzonitrile-sliver shell-goat anti-rabbit immunoglobulin G (Au@4-MBN@Ag@IgG) nanoparticles.
- Utilizing the unique core-molecule-shell structure and silver shell for strong SERS signal generation.
- Employing a sandwich immune response for targeted binding of nanoprobes to hERG ion channels on cell surfaces.
- Detection of the 4-MBN reporter molecule's characteristic Raman peak at 2233 cm⁻¹.
Main Results:
- Achieved strong and stable SERS signals due to the nanoprobe's structure.
- Demonstrated successful targeted binding to hERG ion channels via antibody conjugation.
- Quantified antibody concentration and ion channel protein expression by the intensity of the 4-MBN peak.
- Successfully performed in-situ imaging of hERG ion channel distribution on transfected HEK293 cell surfaces.
Conclusions:
- The developed Au@4-MBN@Ag@IgG nanoparticles serve as effective SERS nanoprobes for ion channel investigation.
- This amplification-free SERS strategy enables precise, in-situ profiling of ion channel distribution at the single-cell level.
- This novel approach offers a promising tool for advancing ion channel research and its applications in diagnostics and drug development.
Abstract:
Research on ion channels and their monoclonal antibodies plays a critical role in drug development and disease diagnosis. The current ion channel researches are often not conducted in the microenvironment for cells survival, which restricts the mechanism study of the links between the cell structure and the ion channel function. In this work, we synthesized gold core-4-mercaptobenzonitrile-sliver shell-goat anti-rabbit immunoglobulin G (Au@4-MBN@Ag@IgG) nanoparticles as surface-enhanced Raman scattering (SERS) nanoprobes for investigating the human ether-a-go-go related gene (hERG) potassium ion channel in cell membranes. This is the first attempt to study ion channels using SERS. Due to the unique core-molecule-shell structure and the silver shell of nanoprobes, strong and stable SERS signal was obtained. With the help of antibodies, the Au@4-MBN@Ag@IgG nanoprobes were captured by hERG antibodies and then bonded with hERG ion channels based on the sandwich immune response. The reporter molecule, 4-MBN, displayed a strong and sharp characteristic peak at 2233 cm-1 in the Raman silent region. The intensity of this peak denoted the concentration of antibodies and the expression of ion channel proteins. We successfully applied this amplification-free method for in-situ imaging the distribution of the hERG ion channel on the transfected HEK293 cell surface at the single-cell level. This hERG ion channel profiling strategy promises a maneuverable tool for ion channel research.
