Real-time full-spectral imaging and affinity measurements from 50 microfluidic channels using nanohole surface plasmon resonance.
With recent advances in high-throughput proteomics and systems biology, there is a growing demand for new instruments that can precisely quantify a wide range of receptor-ligand binding kinetics in a high-throughput fashion. Here we demonstrate a surface plasmon resonance (SPR) imaging spectroscopy instrument capable of simultaneously extracting binding kinetics and affinities from 50 parallel microfluidic channels. The instrument utilizes large-area (~ cm(2)) metallic nanohole arrays as SPR sensing substrates and combines a broadband light source, a high-resolution imaging spectrometer and a low-noise CCD camera to extract spectral information from every channel in real time with a refractive index resolution of 7.7 × 10(-6) refractive index units. To demonstrate the utility of our instrument for quantifying a wide range of biomolecular interactions, each parallel microfluidic channel is coated with a biomimetic supported lipid membrane containing ganglioside (GM1) receptors. The binding kinetics of cholera toxin b (CTX-b) to GM1 are then measured in a single experiment from 50 channels. By combining the highly parallel microfluidic device with large-area periodic nanohole array chips, our SPR imaging spectrometer system enables high-throughput, label-free, real-time SPR biosensing, and its full-spectral imaging capability combined with nanohole arrays could enable integration of SPR imaging with concurrent surface-enhanced Raman spectroscopy.
Laboratory of Nanostructures and Biosensing, Department of Electrical and Computer Engineering, University of Minnesota, Minneapolis, MN 55455, USA.
Si Hoon
SH
Lee
Nathan C
NC
Lindquist
Nathan J
NJ
Wittenberg
Luke R
LR
Jordan
Sang-Hyun
SH
Oh
D040761:Biomimetic Materials / Q000737:chemistry; D002772:Cholera Toxin / Q000737:chemistry*; D005677:G(M1) Ganglioside / Q000737:chemistry*; D008567:Membranes, Artificial; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D020349:Surface Plasmon Resonance / Q000295:instrumentation* / Q000379:methods*
D008567:Membranes, Artificial; D005677:G(M1) Ganglioside; D002772:Cholera Toxin