Measuring the Acoustic Reflex through the Tympanic Membrane.

INTRODUCTION: The acoustic reflex is the active response of the middle ear to loud sounds, altering the mechanical transfer function of the acoustic energy into the inner ear. Our goal was to observe the effect of the acoustic reflex on the tympanic membrane by identifying a significant nonlinear increase in membrane oscillations. METHODS: By using interferometric spectrally encoded endoscopy, we record the membrane oscillations over time in response to a loud, 200-ms-long acoustic stimulus. RESULTS: A gradual reflex activation is measured between approximately 40 and 100 ms, manifested as a linear 42% increase in the umbo oscillation amplitude. CONCLUSION: The measured oscillations correlate well with those expected from a mechanical model of a damped harmonic oscillator, and the results of this work demonstrate the potential of interferometric spectrally encoded endoscopy to observe unique dynamical processes in the tympanic membrane and in the middle ear.

A Triangular-Matrix-Based Spectral Encoding Method for Broadband Filtering and Reconstruction-Based Spectral Measurement.

Broadband filtering and reconstruction-based spectral measurement represent a hot technical route for miniaturized spectral measurement; the measurement encoding scheme has a great effect on the spectral reconstruction fidelity. The existing spectral encoding schemes are usually complex and hard to implement; thus, the applications are severely limited. Considering this, here, a simple spectral encoding method based on a triangular matrix is designed. The condition number of the proposed spectral encoding system is estimated and demonstrated to be relatively low theoretically; then, verification experiments are carried out, and the results show that the proposed encoding can work well under precise or unprecise encoding and measurement conditions; therefore, the proposed scheme is demonstrated to be an effective trade-off of the spectral encoding efficiency and implementation cost.

A Spectral Encoding Simulator for Broadband Active Illumination and Reconstruction-Based Spectral Measurement.

Spectral reflectance or transmittance measurements provide intrinsic information on the material of an object and are widely used in remote sensing, agriculture, diagnostic medicine, etc. Most reconstruction-based spectral reflectance or transmittance measurement methods based on broadband active illumination use narrow-band LEDs or lamps combined with specific filters as spectral encoding light sources. These light sources cannot achieve the designed spectral encoding with a high resolution and accuracy due to their low degree of freedom for adjustment, leading to inaccurate spectral measurements. To address this issue, we designed a spectral encoding simulator for active illumination. The simulator is composed of a prismatic spectral imaging system and a digital micromirror device. The spectral wavelengths and intensity are adjusted by switching the micromirrors. We used it to simulate spectral encodings according to the spectral distribution on micromirrors and solved the DMD patterns corresponding to the spectral encodings with a convex optimization algorithm. To verify the applicability of the simulator for spectral measurements based on active illumination, we used it to numerically simulate existing spectral encodings. We also numerically simulated a high-resolution Gaussian random measurement encoding for compressed sensing and measured the spectral reflectance of one vegetation type and two minerals through numerical simulations. We reconstructed the spectral transmittance of a calibrated filter through an experiment. The results show that the simulator can measure the spectral reflectance or transmittance with a high resolution and accuracy.

Accelerated MR spectroscopic imaging-a review of current and emerging techniques.

Over more than 30 years in vivo MR spectroscopic imaging (MRSI) has undergone an enormous evolution from theoretical concepts in the early 1980s to the robust imaging technique that it is today. The development of both fast and efficient sampling and reconstruction techniques has played a fundamental role in this process. State-of-the-art MRSI has grown from a slow purely phase-encoded acquisition technique to a method that today combines the benefits of different acceleration techniques. These include shortening of repetition times, spatial-spectral encoding, undersampling of k-space and time domain, and use of spatial-spectral prior knowledge in the reconstruction. In this way in vivo MRSI has considerably advanced in terms of spatial coverage, spatial resolution, acquisition speed, artifact suppression, number of detectable metabolites and quantification precision. Acceleration not only has been the enabling factor in high-resolution whole-brain 1 H-MRSI, but today is also common in non-proton MRSI (31 P, 2 H and 13 C) and applied in many different organs. In this process, MRSI techniques had to constantly adapt, but have also benefitted from the significant increase of magnetic field strength boosting the signal-to-noise ratio along with high gradient fidelity and high-density receive arrays. In combination with recent trends in image reconstruction and much improved computation power, these advances led to a number of novel developments with respect to MRSI acceleration. Today MRSI allows for non-invasive and non-ionizing mapping of the spatial distribution of various metabolites' tissue concentrations in animals or humans, is applied for clinical diagnostics and has been established as an important tool for neuro-scientific and metabolism research. This review highlights the developments of the last five years and puts them into the context of earlier MRSI acceleration techniques. In addition to 1 H-MRSI it also includes other relevant nuclei and is not limited to certain body regions or specific applications.

Density-weighted concentric circle trajectories for high resolution brain magnetic resonance spectroscopic imaging at 7T.

PURPOSE: Full-slice magnetic resonance spectroscopic imaging at ≥7 T is especially vulnerable to lipid contaminations arising from regions close to the skull. This contamination can be mitigated by improving the point spread function via higher spatial resolution sampling and k-space filtering, but this prolongs scan times and reduces the signal-to-noise ratio (SNR) efficiency. Currently applied parallel imaging methods accelerate magnetic resonance spectroscopic imaging scans at 7T, but increase lipid artifacts and lower SNR-efficiency further. In this study, we propose an SNR-efficient spatial-spectral sampling scheme using concentric circle echo planar trajectories (CONCEPT), which was adapted to intrinsically acquire a Hamming-weighted k-space, thus termed density-weighted-CONCEPT. This minimizes voxel bleeding, while preserving an optimal SNR. THEORY AND METHODS: Trajectories were theoretically derived and verified in phantoms as well as in the human brain via measurements of five volunteers (single-slice, field-of-view 220 × 220 mm2 , matrix 64 × 64, scan time 6 min) with free induction decay magnetic resonance spectroscopic imaging. Density-weighted-CONCEPT was compared to (a) the originally proposed CONCEPT with equidistant circles (here termed e-CONCEPT), (b) elliptical phase-encoding, and (c) 5-fold Controlled Aliasing In Parallel Imaging Results IN Higher Acceleration accelerated elliptical phase-encoding. RESULTS: By intrinsically sampling a Hamming-weighted k-space, density-weighted-CONCEPT removed Gibbs-ringing artifacts and had in vivo +9.5%, +24.4%, and +39.7% higher SNR than e-CONCEPT, elliptical phase-encoding, and the Controlled Aliasing In Parallel Imaging Results IN Higher Acceleration accelerated elliptical phase-encoding (all P < 0.05), respectively, which lead to improved metabolic maps. CONCLUSION: Density-weighted-CONCEPT provides clinically attractive full-slice high-resolution magnetic resonance spectroscopic imaging with optimal SNR at 7T. Magn Reson Med 79:2874-2885, 2018. © 2017 The Authors Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Profiling DNA mutation patterns by SERS fingerprinting for supervised cancer classification.

Profiling DNA mutation patterns for cancer classification plays an essential role in precision and personalized medicine. Conventional PCR-based mutation assay is limited by the extensive labour on target amplification. We herein create an amplification-free surface enhanced Raman spectroscopy (SERS) biochip which enables direct and simultaneous identification of multiple point mutations in tumor cells. Without pre-amplifying the target sequences, the SERS assay reads out the presence of cellular mutations through the interpretation of Raman fingerprints. The SERS sensor is integrated into a microfluidic chip, achieving one-step multiplex analysis within 40 min. Importantly, by combining SERS spectra encoding technique with supervised learning algorithm, a panel of nucleotide mixtures can be well distinguished according to their mutation profiles. We initially demonstrate an excellent levels of classification in samples from colorectal cancer and melanoma cell lines. For final clinical validation, the system performance is verified by classifying cancer patient samples, which shows an accuracy above 90%. Due to the simplicity and rapidness, the SERS biosensor is expected to become a promising tool for clinical point-of-care diagnosis towards precision medicine.

Protocol for Peptide Synthesis on Spectrally Encoded Beads for MRBLE-pep Assays.

Every living cell relies on signal transduction pathways comprised of protein-protein interactions (PPIs). In many cases, these PPIs are between a folded protein domain and a short linear motif (SLiM) within an unstructured region of a protein. As a result of this small interaction interface (3-10 amino acids), the affinities of SLiM-mediated interactions are typically weak (K ds of ~1-10 µM), allowing physiologically relevant changes in cellular concentrations of either protein partner to dictate changes in occupancy and thereby transmit cellular signals. However, these weak affinities also render detection and quantitative measurement of these interactions challenging and labor intensive. To address this, we recently developed MRBLE-pep, a technology that employs peptide libraries synthesized on spectrally encoded hydrogel beads to allow multiplexed affinity measurements between a protein and many different peptides in parallel. This approach dramatically reduces both the amount of protein and peptide as well as the time required to measure protein-peptide affinities compared to traditional methods. Here, we provide a detailed protocol describing how to: (1) functionalize polyethylene glycol diacrylate (PEG-DA) MRBLE beads with free amine groups, (2) synthesize peptide libraries on functionalized MRBLEs, (3) validate synthesized peptide sequences via MALDI mass spectrometry and quantify evenness of peptide coverage on MRBLEs, (4) use MRBLE-bound peptide libraries in multiplexed protein binding assays, and (5) analyze binding data to determine binding affinities. We anticipate that this protocol should prove useful for other researchers seeking to use MRBLE-pep in their own laboratories as well as for researchers broadly interested in solid-phase peptide synthesis and protein-protein binding assay development.

Quantitative mapping of protein-peptide affinity landscapes using spectrally encoded beads.

Transient, regulated binding of globular protein domains to Short Linear Motifs (SLiMs) in disordered regions of other proteins drives cellular signaling. Mapping the energy landscapes of these interactions is essential for deciphering and perturbing signaling networks but is challenging due to their weak affinities. We present a powerful technology (MRBLE-pep) that simultaneously quantifies protein binding to a library of peptides directly synthesized on beads containing unique spectral codes. Using MRBLE-pep, we systematically probe binding of calcineurin (CN), a conserved protein phosphatase essential for the immune response and target of immunosuppressants, to the PxIxIT SLiM. We discover that flanking residues and post-translational modifications critically contribute to PxIxIT-CN affinity and identify CN-binding peptides based on multiple scaffolds with a wide range of affinities. The quantitative biophysical data provided by this approach will improve computational modeling efforts, elucidate a broad range of weak protein-SLiM interactions, and revolutionize our understanding of signaling networks.

Programmable Microfluidic Synthesis of Over One Thousand Uniquely Identifiable Spectral Codes.

Encoded microparticles have become a powerful tool for a wide array of applications, including high-throughput sample tracking and massively parallel biological multiplexing. Spectral encoding, where particles are encoded with distinct luminescence spectra, provides a particularly appealing encoding strategy because of the ease of reading codes and assay flexibility. To date, spectral encoding has been limited in the number of codes that can be accurately resolved. Here, we demonstrate an automated 5-dimensional spectral encoding scheme using lanthanide nanophosphors that is capable of producing isotropic spherical microparticles with up to 1,100 unique codes, which we term MRBLEs (Microspheres with Ratiometric Barcode Lanthanide Encoding). We further develop a quantitative framework for evaluating global ability to distinguish codes and demonstrate that for six different sets of MRBLEs ranging from 106 to 1,101 codes in size, > 98% of MRBLEs can be assigned to a code with 99.99% confidence. These > 1,000 code sets represent the largest spectral code libraries built to date. We expect that these MRBLEs will enable a wide variety of novel multiplexed assays.