title: Bioinspired Protein Channel-Based Scanning Ion Conductance Microscopy (Bio-SICM) for Simultaneous Conductance and Specific Molecular Imaging.
abstract: The utility of stochastic single-molecule detection using protein nanopores has found widespread application in bioanalytical sensing as a result of the inherent signal amplification of the resistive pulse method. Integration of protein nanopores with high-resolution scanning ion conductance microscopy (SICM) extends the utility of SICM by enabling selective chemical imaging of specific target molecules, while simultaneously providing topographical information about the net ion flux through a pore under a concentration gradient. In this study, we describe the development of a bioinspired scanning ion conductance microscopy (bio-SICM) approach that couples the imaging ability of SICM with the sensitivity and chemical selectivity of protein channels to perform simultaneous pore imaging and specific molecule mapping. To establish the framework of the bio-SICM platform, we utilize the well-studied protein channel α-hemolysin (αHL) to map the presence of β-cyclodextrin (βCD) at a substrate pore opening. We demonstrate concurrent pore and specific molecule imaging by raster scanning an αHL-based probe over a glass membrane containing a single 25-μm-diameter glass pore while recording the lateral positions of the probe and channel activity via ionic current. We use the average channel current to create a conductance image and the raw current-time traces to determine spatial localization of βCD. With further optimization, we believe that the bio-SICM platform will provide a powerful analytical methodology that is generalizable, and thus offers significant utility in a myriad of bioanalytical applications.
authors: Department of Chemistry and Biochemistry, University of Maryland Baltimore County , Baltimore, Maryland 21250, United States.
authors: Florika C
authors: FC
authors: Macazo
authors: Department of Chemistry and Biochemistry, University of Maryland Baltimore County , Baltimore, Maryland 21250, United States.
authors: Ryan J
authors: RJ
authors: White
mesh_terms: D029968:Escherichia coli Proteins / Q000737:chemistry*; D006460:Hemolysin Proteins / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry; D020527:Microscopy, Scanning Probe / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D013269:Stochastic Processes
chemical_list: D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D008051:Lipid Bilayers