Spatially Resolved Protein Binding Kinetics Analysis in Microfluidic Photonic Crystal Sensors.

Organ-on-a-Chip systems are emerging as an important in vitro analysis method for drug screening and medical research. For continuous biomolecular monitoring of the cell culture response, label-free detection within the microfluidic system or in the drainage tube is promising. We study photonic crystal slabs integrated with a microfluidic chip as an optical transducer for label-free biomarker detection with a non-contact readout of binding kinetics. This work analyzes the capability of same-channel reference for protein binding measurements by using a spectrometer and 1D spatially resolved data evaluation with a spatial resolution of 1.2 µm. A cross-correlation-based data-analysis procedure is implemented. First, an ethanol-water dilution series is used to obtain the limit of detection (LOD). The median of all row LODs is (2.3±0.4)×10-4 RIU with 10 s exposure time per image and (1.3±0.24)×10-4 RIU with 30 s exposure time. Next, we used a streptavidin-biotin binding process as a test system for binding kinetics. Time series of optical spectra were recorded while constantly injecting streptavidin in DPBS at concentrations of 1.6 nM, 3.3 nM, 16.6 nM and 33.3 nM into one channel half as well as the whole channel. The results show that localized binding within a microfluidic channel is achieved under laminar flow. Furthermore, binding kinetics are fading out at the microfluidic channel edge due to the velocity profile.

Intensity-Based Camera Setup for Refractometric and Biomolecular Sensing with a Photonic Crystal Microfluidic Chip.

Label-free sensing is a promising approach for point-of-care testing devices. Among optical transducers, photonic crystal slabs (PCSs) have positioned themselves as an inexpensive yet versatile platform for label-free biosensing. A spectral resonance shift is observed upon biomolecular binding to the functionalized surface. Commonly, a PCS is read out by a spectrometer. Alternatively, the spectral shift may be translated into an intensity change by tailoring the system response. Intensity-based camera setups (IBCS) are of interest as they mitigate the need for postprocessing, enable spatial sampling, and have moderate hardware requirements. However, they exhibit modest performance compared with spectrometric approaches. Here, we show an increase of the sensitivity and limit of detection (LOD) of an IBCS by employing a sharp-edged cut-off filter to optimize the system response. We report an increase of the LOD from (7.1 ± 1.3) × 10-4 RIU to (3.2 ± 0.7) × 10-5 RIU. We discuss the influence of the region of interest (ROI) size on the achievable LOD. We fabricated a biochip by combining a microfluidic and a PCS and demonstrated autonomous transport. We analyzed the performance via refractive index steps and the biosensing ability via diluted glutathione S-transferase (GST) antibodies (1:250). In addition, we illustrate the speed of detection and demonstrate the advantage of the additional spatial information by detecting streptavidin (2.9 µg/mL). Finally, we present the detection of immunoglobulin G (IgG) from whole blood as a possible basis for point-of-care devices.

Near cut-off wavelength operation of resonant waveguide grating biosensors.

Numerical simulations and analytical calculations are performed to support the design of grating-coupled planar optical waveguides for biological sensing. Near cut-off and far from cut-off modes are investigated, and their characteristics and suitability for sensing are compared. The numerical simulations reveal the high sensitivity of the guided mode intensity near the cut-off wavelength for any refractive index change along the waveguide. Consequently, it is sufficient to monitor the intensity change of the near cut-off sensing mode, which leads to a simpler sensor design compared to those setups where the resonant wavelength shift of the guided mode is monitored with high precision. The operating wavelength and the sensitivity of the proposed device can be tuned by varying the geometrical parameters of the corrugated waveguide. These results may lead to the development of highly sensitive integrated sensors, which have a simple design and therefore are cost-effective for a wide range of applications. These numerical findings are supported with experimental results, where the cut-off sensing mode was identified.