Critical angle reflection imaging for quantification of molecular interactions on glass surface
Data files
Apr 27, 2021 version files 12.42 GB
-
ACh-nAChR_binding_in_cells_(Fig5).zip
5.04 GB
-
Biomolecular_interactions_(Fig2).xlsx
497.60 KB
-
CAR_response_to_ethanol_(Fig1).xlsx
84.15 KB
-
Small_molecule_detection_(Fig3).xlsx
182.06 KB
-
Supplementary_Information.zip
427.39 MB
-
WGA-glycoprotein_binding_in_cells_(Fig4).zip
6.95 GB
Abstract
Quantification of molecular interactions on a surface is typically achieved via label-free techniques such as surface plasmon resonance (SPR). The sensitivity of SPR originates from the characteristic that the SPR angle is sensitive to the surface refractive index change. Analogously, in another interfacial optical phenomenon, total internal reflection, the critical angle is also refractive index dependent. Therefore, surface refractive index change can also be quantified by measuring the reflectivity near critical angle. Based on this concept, we develop a method called critical angle reflection (CAR) imaging to quantify molecular interactions on glass surface. CAR imaging can be performed on SPR imaging setups. Through a side-by-side comparison, we show that CAR is capable of most molecular interaction measurements that SPR does, including proteins, nucleic acids and cell-based detections. In addition, we show that CAR can detect small molecule bindings and intracellular signals beyond SPR sensing range. CAR exhibits several distinct characteristics over SPR, including tunable sensitivity and dynamic range, deeper vertical sensing range, fluorescence compatibility, broader wavelength and polarization of light selection, and glass surface chemistry. We anticipate CAR can expand SPR capability in small molecule detection, whole cell-based detection, simultaneous fluorescence imaging, and broader conjugation chemistry.
The dataset include binding kinetics data measured by CAR and SPR imaging techniques.
Please use Winspall to open WSP file, and use Fiji (ImageJ) to open image files.