Distributed interferometric fiber tip biosensors for a multi-channel and label-free biomolecular interaction analysis.

This paper describes fabrication and implementation of distributed optical fiber tip biosensor probes for simultaneously measuring label-free biomolecular interactions at multiple locations. Biosensor probes at the tip of a single-mode fiber are Fabry-Perot etalons that are functionalized with a capture layer for a specific biomolecule. A coherence multiplexing method is implemented to separate data collected from distributed biosensors in a single data stream. Multiplexing is achieved by using fiber tip biosensors of varying etalon lengths with the same or different capture layers for each biosensing channel. Experiments demonstrating simultaneous multi-channel recording of protein-to-protein interaction sensorgrams with fiber tip biosensor probes are presented.

Gliomas Interact with Non-glioma Brain Cells via Extracellular Vesicles.

Emerging evidence suggests that crosstalk between glioma cells and the brain microenvironment may influence brain tumor growth. To date, known reciprocal interactions among these cells have been limited to the release of paracrine factors. Combining a genetic strategy with longitudinal live imaging, we find that individual gliomas communicate with distinct sets of non-glioma cells, including glial cells, neurons, and vascular cells. Transfer of genetic material is achieved mainly through extracellular vesicles (EVs), although cell fusion also plays a minor role. We further demonstrate that EV-mediated communication leads to the increase of synaptic activity in neurons. Blocking EV release causes a reduction of glioma growth in vivo. Our findings indicate that EV-mediated interaction between glioma cells and non-glioma brain cells alters the tumor microenvironment and contributes to glioma development.

Label-free optical detection of action potential in mammalian neurons.

We describe an optical technique for label-free detection of the action potential in cultured mammalian neurons. Induced morphological changes due to action potential propagation in neurons are optically interrogated with a phase sensitive interferometric technique. Optical recordings composed of signal pulses mirror the electrical spike train activity of individual neurons in a network. The optical pulses are transient nanoscale oscillatory changes in the optical path length of varying peak magnitude and temporal width. Exogenous application of glutamate to cortical neuronal cultures produced coincident increase in the electrical and optical activity; both were blocked by application of a Na-channel blocker, Tetrodotoxin. The observed transient change in optical path length in a single optical pulse is primarily due to physical fluctuations of the neuronal cell membrane mediated by a yet unknown electromechanical transduction phenomenon. Our analysis suggests a traveling surface wave in the neuronal cell membrane is responsible for the measured optical signal pulses.

Probabilistic partial least squares regression for quantitative analysis of Raman spectra.

With the latest development of Surface-Enhanced Raman Scattering (SERS) technique, quantitative analysis of Raman spectra has shown the potential and promising trend of development in vivo molecular imaging. Partial Least Squares Regression (PLSR) is state-of-the-art method. But it only relies on training samples, which makes it difficult to incorporate complex domain knowledge. Based on probabilistic Principal Component Analysis (PCA) and probabilistic curve fitting idea, we propose a probabilistic PLSR (PPLSR) model and an Estimation Maximisation (EM) algorithm for estimating parameters. This model explains PLSR from a probabilistic viewpoint, describes its essential meaning and provides a foundation to develop future Bayesian nonparametrics models. Two real Raman spectra datasets were used to evaluate this model, and experimental results show its effectiveness.

Models and methods for quantitative analysis of surface-enhanced Raman spectra.

The quantitative analysis of surface-enhanced Raman spectra using scattering nanoparticles has shown the potential and promising applications in in vivo molecular imaging. The diverse approaches have been used for quantitative analysis of Raman pectra information, which can be categorized as direct classical least squares models, full spectrum multivariate calibration models, selected multivariate calibration models, and latent variable regression (LVR) models. However, the working principle of these methods in the Raman spectra application remains poorly understood and a clear picture of the overall performance of each model is missing. Based on the characteristics of the Raman spectra, in this paper, we first provide the theoretical foundation of the aforementioned commonly used models and show why the LVR models are more suitable for quantitative analysis of the Raman spectra. Then, we demonstrate the fundamental connections and differences between different LVR methods, such as principal component regression, reduced-rank regression, partial least square regression (PLSR), canonical correlation regression, and robust canonical analysis, by comparing their objective functions and constraints.We further prove that PLSR is literally a blend of multivariate calibration and feature extraction model that relates concentrations of nanotags to spectrum intensity. These features (a.k.a. latent variables) satisfy two purposes: the best representation of the predictor matrix and correlation with the response matrix. These illustrations give a new understanding of the traditional PLSR and explain why PLSR exceeds other methods in quantitative analysis of the Raman spectra problem. In the end, all the methods are tested on the Raman spectra datasets with different evaluation criteria to evaluate their performance.

Continuous wavelet transform based partial least squares regression for quantitative analysis of Raman spectrum.

Quantitative analysis of Raman spectra using surface-enhanced Raman scattering (SERS) nanoparticles has shown the potential and promising trend of development in in vivo molecular imaging. Partial least square regression (PLSR) methods have been reported as state-of-the-art methods. However, the approaches fully rely on the intensities of Raman spectra and can not avoid the influences of the unstable background. In this paper we design a new continuous wavelet transform based PLSR (CWT-PLSR) algorithm that uses mixing concentrations and the average CWT coefficients of Raman spectra to carry out PLSR. We elaborate and prove how the average CWT coefficients with a Mexican hat mother wavelet are robust representations of Raman peaks, and the method can reduce the influences of unstable baseline and random noises during the prediction process. The algorithm was tested using three Raman spectra data sets with three cross-validation methods in comparison with current leading methods, and the results show its robustness and effectiveness.

Eigenspectra, a robust regression method for multiplexed Raman spectra analysis.

With the latest development of Surface Enhanced Raman Scattering (SERS) nanoparticles, Raman spectroscopy now can be extended to bioimaging and biosensing. In this study, we demonstrate the ability of Raman spectroscopy to separate multiple spectral fingerprints using Raman nanotags. A machine learning method is proposed to estimate the mixing ratios of sources from mixture signals. It decomposes the mixture signals into components for both best representation and most relating to mixing ratios. Then regression coefficients are calculated for the prediction. The robustness of the method was compared with least squares and weighted least squares methods.

Coherence-multiplexed, label-free biomolecular interaction analysis.

This Letter describes an interferometric technique based on the principle of coherence multiplexing for multichannel, label-free biosensing applications. Multiple biosensors can be interrogated simultaneously with a single spectral-domain, phase-sensitive interferometer by coding the individual sensograms in coherence-multiplexed channels. The experimental results demonstrate the multiplexed quantitative biomolecular interaction of antibodies binding to antigen-coated functionalized biosensor chip surfaces. The described technique also applies to a variety of other distributed and multiplexed sensing applications besides biosensing.

Real-time detection of implant-associated neutrophil responses using a formyl peptide receptor-targeting NIR nanoprobe.

Neutrophils play an important role in implant-mediated inflammation and infection. Unfortunately, current methods which monitor neutrophil activity, including enzyme measurements and histological evaluation, require many animals and cannot be used to accurately depict the dynamic cellular responses. To understand the neutrophil interactions around implant-mediated inflammation and infection it is critical to develop methods which can monitor in vivo cellular activity in real time. In this study, formyl peptide receptor (FPR)-targeting near-infrared nanoprobes were fabricated. This was accomplished by conjugating near-infrared dye with specific peptides having a high affinity to the FPRs present on activated neutrophils. The ability of FPR-targeting nanoprobes to detect and quantify activated neutrophils was assessed both in vitro and in vivo. As expected, FPR-targeting nanoprobes preferentially accumulated on activated neutrophils in vitro. Following transplantation, FPR-targeting nanoprobes preferentially accumulated at the biomaterial implantation site. Equally important, a strong relationship was observed between the extent of fluorescence intensity in vivo and the number of recruited neutrophils at the implantation site. Furthermore, FPR-targeting nanoprobes may be used to detect and quantify the number of neutrophils responding to a catheter-associated infection. The results show that FPR-targeting nanoprobes may serve as a powerful tool to monitor and measure the extent of neutrophil responses to biomaterial implants in vivo.

Gold nanotags for combined multi-colored Raman spectroscopy and x-ray computed tomography.

Multi-color gold-nanoparticle-based tags (nanotags) are synthesized for combined surface-enhanced Raman spectroscopy (SERS) and x-ray computed tomography (CT). The nanotags are synthesized with quasi-spherical gold nanoparticles encoded with a reporter dye (color), each with a unique Raman spectrum. A library of nanotags with six different colors were synthesized for a range of gold nanoparticle sizes and an optimum size has been established to yield the largest SERS intensity and x-ray attenuation that is higher than the iodinated CT contrast agents used in clinics. Proof-of-principle in vivo imaging results with nanotags are presented that, for the first time, demonstrates the combined in vivo dual modality imaging capability of SERS and CT with a single nanoparticle probe.

Multi-color Raman nanotags for tumor cell biomarker detection.

We report the synthesis and characterization of multi-color gold nanoparticle based tags (Raman Nanotags) for molecular imaging. The multi-color Raman Nanotags are PEGylated gold nanoparticles (AuNPs) encapsulating a Raman reporter dyes which can be functionalized with any ligand of interest for targeted molecular imaging. The Raman Nanotags synthesized with 65 nm gold nanoparticles exhibit the largest surface enhanced Raman scattering (SERS) signal. Results are presented quantify the measured SERS signal, dynamic range, reproducibility, and stability of Raman Nanotags. In vitro cell culture experiments for targeted biomarker detection using functionalized Raman Nanotags are also presented.

Neuro-optical microfluidic platform to study injury and regeneration of single axons.

We describe a neuro-optical microfluidic platform for studying injury and subsequent regeneration of individual mammalian axons. This platform consists of three components integrated on an inverted microscope, which include a compartmentalized neuronal culture microfluidic device, a femtosecond laser to enable precise axotomy, and a custom built mini cell culture incubator for continuous long term observation of post injury events. We demonstrate the unique capabilities of the platform by injuring individual central and peripheral nervous system axons and monitoring the post injury sequence of events from initial degeneration to subsequent regeneration. This platform will enable study and understanding of neuronal response to injury that is currently not possible with conventional cell culture platform and tools.

Phase sensitive interferometry for biosensing applications.

A simple yet highly sensitive implementation of an interferometric technique for a label-free molecular biosensing application is described. The intereferometric detection method is based on the phase-sensitive detection of spectral interference fringes. The change in optical path length due to binding of biomolecules on functionalized optically clear substrates can be quantified by detecting the change in the phase of the spectral fringes. The common path interferometeric design permits measurement of sub-monolayer binding of biomolecules to the sensor surfaces.

Spatial refractive index measurement of porcine artery using differential phase optical coherence microscopy.

BACKGROUND AND OBJECTIVES: We describe a methodology to record spatial variation of refractive index of porcine renal artery using differential phase optical coherence microscopy (DP-OCM). STUDY DESIGN/MATERIALS AND METHODS: The DP-OCM provides quantitative measurement of thin specimen phase retardation and refractive index by measuring optical path-length changes on the order of a few nanometers and with a lateral resolution of 3 microm. The DP-OCM instrumentation is an all-fiber, dual-channel Michelson interferometer constructed using a polarization maintaining (PM) fiber. RESULTS: Two-dimensional en face dual-channel phase images are taken over a 150 x 200 microm region on a microscopic slide, and the images are reconstructed by plotting a two-dimensional refractive index map as the OCM beam is moved across the sample. CONCLUSIONS: Because the DP-OCM can record transient changes in the optical path-length, the system may be used to record quantitative optical path-length alterations of tissue in response to various stimuli. A fiber-based DP-OCM may have the potential to substantially improve in vivo imaging of individual cells for a variety of clinical diagnostics, and monitoring applications.

Quantitative phase-contrast imaging of cells with phase-sensitive optical coherence microscopy.

We describe a method for en face phase-contrast imaging of cells with a fiber-based differential phase-contrast optical coherence microscopy system. Recorded en face images are quantitative phase-contrast maps of cells due to spatial variation of the refractive index and (or) thickness of various cellular components. Quantitative phase-contrast images of human epithelial cheek cells obtained with the fiber-based differential phase-contrast optical coherence microscopy system are presented.

Differential phase optical coherence probe for depth-resolved detection of photothermal response in tissue.

We describe a differential phase low-coherence interferometric probe for non-invasive, quantitative imaging of photothermal phenomena in biological materials. Our detection method utilizes principles of optical coherence tomography with differential phase measurement of interference fringe signals. A dual-channel optical low-coherence probe is used to analyse laser-induced thermoelastic and thermorefractive effects in tissue with micrometre axial resolution and nanometre sensitivity. We demonstrate an application of the technique using tissue phantoms and ex-vivo tissue specimens of rodent dorsal skin.

Imaging tissue response to electrical and photothermal stimulation with nanometer sensitivity.

BACKGROUND AND OBJECTIVES: Tissue response to thermal, electrical, or chemical stimuli are important in the health and survival of tissue. We report experimental results to assess tissue response to various stimuli using a low coherence differential phase interferometer. STUDY DESIGN/MATERIALS AND METHODS: The optical system utilized to measure tissue response is a novel fiber-based phase sensitive optical low coherence reflectometer (PS-OLCR). Inasmuch as the PS-OLCR works with back-reflected light, noninvasive sensing of tissue response to stimuli is possible. In addition to high lateral (approximately 10 microm) and longitudinal (approximately 10 microm) resolution, PS-OLCR can measure sub-wavelength changes in optical path-length (Angstrom/nanometer range) by extracting the phase difference between interference fringes in two channels corresponding to orthogonal polarization modes. RESULTS: When light spatially splits into two polarization states, precise analysis of surface topography or tissue surface response such as swelling or collapse are possible. Time resolved measurements of nanometer-scale path length changes in response to electrical and thermal stimuli are demonstrated using longitudinally delayed polarization channels. CONCLUSIONS: Since PS-OLCR is a useful tool to detect ultra-small path length changes, the system has potential to aid scientists in investigating important phenomena in biomaterials and developing useful diagnostic and therapeutic imaging modalities. Applications include tissue surface profilometry, measurement of tissue, and cell response to various stimuli, high-resolution intensity and phase imaging.

Polarization-maintaining fiber-based optical low-coherence reflectometer for characterization and ranging of birefringence.

We describe a polarization-maintaining fiber-based polarization-sensitive optical low-coherence reflectometer for measurement of depth-resolved birefringence. Unlike for other fiber-based polarization-sensitive optical low-coherence reflectometers, here the linear birefringence of a sample can be measured from data recorded in a single A scan. Simultaneous measurement of retardation and orientation of birefringent axes with mica wave plates is demonstrated. The measured retardation is insensitive to sample rotation in the plane perpendicular to ranging.

Refractive-index profiling of embedded microstructures in optical materials.

We describe use of a phase-sensitive low-coherence reflectometer to measure spatial variation of refractive index in optical materials. The described interferometric technique is demonstrated to be a valuable tool to profile the refractive index of optical elements such as integrated waveguides and photowritten optical microstructures. As an example, a refractive-index profile is mapped of a microstructure written in a microscope glass slide with an ultrashort-pulse laser.