High-speed lymphocyte tracking provided the spatial and temporal resolution to measure lymphocyte flux throughout the phasic contraction. Poiseuille flow was a reasonable model for estimating wall shear stress through most of the phasic contraction cycle of the intervalvular lymphatic segments. Shear rate was low but had large variations in magnitude compared to that seen in blood vessels.
A fluorescence biosensor is described that is based on a photopolymerized poly(ethylene glycol) (PEG) hydrogel incorporating fluorescein isothiocyanate dextran (FITC-dextran) and tetramethylrhodamine isothiocyanate concanavalin A (TRITC-Con A) chemically conjugated into the hydrogel network using an alpha-acryloyl, omega-N-hydroxysuccinimidyl ester of PEG-propionic acid. In the absence of glucose, TRITC-Con A binds with FITC-dextran, and the FITC fluorescence is quenched through fluorescence resonance energy transfer. Competitive glucose binding to TRITC-Con A liberates FITC-dextran, resulting in increased FITC fluorescence proportional to the glucose concentration. In vitro experiments of hydrogel spheres in a solution of 0.1 M phosphate-buffered saline (pH 7.2) and glucose were conducted for multiple TRITC-Con A/FITC-dextran ratios. Hydrogels were characterized on the basis of the percent change in fluorescence intensity when FITC-dextran was liberated by increasing glucose concentrations. The optimum fluorescent change between 0 and 800 mg/dL was obtained with a TRITC-Con A/FITC-dextran mass ratio of 500:5 micrograms/mL PEG. Fluorescent response was linear up to 600 mg/dL. At higher concentrations, the response saturated due to the displacement of the majority of the FITC-dextran and to concentration quenching by free FITC-dextran. Dynamic fluorescent change upon glucose addition was approximately 10 min for a glucose concentration step change from 0 to 200 mg/dL.
Recent technological advancements in the photonics industry have led to a resurgence of interest in optical glucose sensing and to realistic progress toward the development of an optical glucose sensor. Such a sensor has the potential to significantly improve the quality of life for the estimated 16 million diabetics in this country by making routine glucose measurements more convenient. Currently over 100 small companies and universities are working to develop noninvasive or minimally invasive glucose sensing technologies, and optical methods play a large role in these efforts. This article reviews many of the recent advances in optical glucose sensing including optical absorption spectroscopy, polarimetry, Raman spectroscopy, and fluorescent glucose sensing. In addition a review of calibration and data processing methods useful for optical techniques is presented.
Trace detection of the conformational transition of beta-amyloid peptide (Abeta) from a predominantly alpha-helical structure to beta-sheet could have a large impact in understanding and diagnosing Alzheimer's disease. We demonstrate how a novel nanofluidic biosensor using a controlled, reproducible surface enhanced Raman spectroscopy active site was developed to observe Abeta in different conformational states during the Abeta self-assembly process as well as to distinguish Abeta from confounder proteins commonly found in cerebral spinal fluid.
We present both experimental measurements and Monte-Carlo-based simulations of the diffusely backscattered intensity patterns that arise from illuminating a turbid medium with a polarized laser beam. It is rigorously shown that, because of axial symmetry of the system, only seven elements of the effective backscattering Mueller matrix are independent. A new numerical method that allows simultaneous calculation of all 16 elements of the two-dimensional Mueller matrix is used. To validate our method we compared calculations to measurements from a turbid medium that consisted of polystyrene spheres of different sizes and concentrations in deionized water. The experimental and numerical results are in excellent agreement.
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