Biomedical subjects

Gerard L Coté

Publications and source records attributed to Gerard L Coté.

13 recordsLinked to original sources

PubMed2006

Image correlation algorithm for measuring lymphocyte velocity and diameter changes in contracting microlymphatics.

Efforts have recently been made to estimate wall shear stress throughout the contractile cycle of mesenteric rat lymphatics with a high speed video microscopy system. This was prompted by reports in the literature that lymphatic pumping is related to wall shear stress. While one can estimate wall shear stress by tracking lymphocyte velocity, it is prohibitively tedious to manually track particles over a reasonable time frame for a good number of experiments. To overcome this, an image correlation method similar to digital particle imaging velocimetry was developed and tested on contracting lymphatics to measure both vessel diameter and fluid velocity. The program tracked temporal fluctuations in spatially averaged velocity with a standard error of prediction of 0.4 mm/s. From these studies we have measured velocities ranging from -2 to 4 mm/s. Diameter changes were also measured with a standard error of 7 microm. These algorithms and techniques could be beneficial for investigating various changes in contractile behavior as a function of changes in velocity and wall shear stress.

Algorithms↗

PubMed2006

Microporated PEG spheres for fluorescent analyte detection.

Poly(ethylene glycol) (PEG) hydrogels have been used to encapsulate fluorescently labeled molecules in order to detect a variety of analytes. The hydrogels are designed with a mesh size that will retain the sensing elements while allowing for efficient diffusion of small analytes. Some sensing assays, however, require a conformational change or binding of large macromolecules, which may be sterically prohibited in a dense polymer matrix. A process of hydrogel microporation has been developed to create cavities within PEG microspheres to contain the assay components in solution. This arrangement provides improved motility for large sensing elements, while limiting leaching and increasing sensor lifetime. Three hydrogel compositions, 100% PEG, 50% PEG, and microporated 100% PEG, were used to create pH-sensitive microspheres that were tested for response time and stability. In order to assess motility, a second, more complex sensor, namely a FITC-dextran/TRITC-Con A glucose-specific assay was encapsulated within the microspheres.

Capsules↗

PubMed2005

Competitive binding assay for glucose based on glycodendrimer-fluorophore conjugates.

A new fluorescent glucose assay has been created using Alexa Fluor 647-labeled concanavalin A (Con A) and a fourth-generation PAMAM Alexa Fluor 594-labeled glycodendrimer. This assay has been shown to have a large response to glucose within the biological range and to be capable of functioning within a polymer hydrogel. In this paper, the glucose response is shown to be a single fluorophore-based quenching reaction. Data showing that the sensor is fully reversible and specific through competitive binding between the dendrimer and glucose with Con A are presented. Overall, the assay is shown to have potential over the traditional dextran-based assay because it has a larger dynamic response to physiological glucose concentrations, incorporates longer wavelength dyes that improve signal penetration through dermal tissue, and provides an internal reference in the form of a nonreactive fluorescent label.

Binding, Competitive↗

PubMed2005

Processing of pulse oximeter data using discrete wavelet analysis.

A wavelet-based signal processing technique was employed to improve an implantable blood perfusion monitoring system. Data was acquired from both in vitro and in vivo sources: a perfusion model and the proximal jejunum of an adult pig. Results showed that wavelet analysis could isolate perfusion signals from raw, periodic, in vitro data as well as fast Fourier transform (FFT) methods. However, for the quasi-periodic in vivo data segments, wavelet analysis provided more consistent results than the FFT analysis for data segments of 50, 10, and 5 s in length. Wavelet analysis has thus been shown to require less data points for quasi-periodic data than FFT analysis making it a good choice for an indwelling perfusion monitor where power consumption and reaction time are paramount.

Algorithms↗

PubMed2005

An autocorrelation-based time domain analysis technique for monitoring perfusion and oxygenation in transplanted organs.

In designing an implantable sensor for perfusion monitoring of transplant organs the ability of the sensor to gather perfusion information with limited power consumption and in near real time is paramount. The following work was performed to provide a processing method that is able to predict perfusion and oxygenation change within the blood flowing through a transplanted organ. For this application, an autocorrelation-based algorithm was used to reduce the acquisition time required for fast Fourier transform (FFT) analysis while retaining the accuracy inherent to FFT analysis. In order to provide data proving that the developed method is able to predict perfusion as accurately as FFT two experiments were developed isolating both periodic and quasi-periodic cardiac frequencies. It was shown that the autocorrelation-based method was able to perform comparably with FFT (limited to a sampling frequency of 300 Hz) and maintain accuracy down to acquisition times as low as 4 s in length.

Algorithms↗

PubMed2005

Real-time separation of perfusion and oxygenation signals for an implantable sensor using adaptive filtering.

In this paper, an adaptive filtering algorithm to separate signals due to perfusion and oxygenation has been developed using an 810-nm source, in addition to 660-nm and 940-nm sources, as an internal reference due to its limited oxygen sensitivity. The newly developed algorithm was tested using Monte Carlo simulated data to prove the effectiveness of the 810-nm reference and adaptive algorithm. Following the simulation, an in vitro model was developed to test the algorithm that used a blood flow through system wrapped with tissue. The system had the ability to isolate the effects of perfusion and oxygenation and the algorithm accurately captured the changes in these signals with reliable consistency. Using the serosal surface of the swine jejunum, in vivo data was also taken to analyze the algorithms response to fluctuating perfusion levels like that seen in hemorrhaging or failing transplants. The algorithm was able to extract the perfusion information from the oxygenation information in this in vivo study. Overall, it was shown that an adaptive filtering algorithm using an 810-nm reference has provided a means to separate oxygenation and perfusion.

Algorithms↗

PubMed2003

Feasibility study using surface-enhanced Raman spectroscopy for the quantitative detection of excitatory amino acids.

The release of excitatory amino acids (EAAs) from injured neurons has been associated with secondary injury following head trauma. The development of a rapid and sensitive method for the quantification of EAAs may provide a means for clinical management of patients affected by head trauma. We explore the potential application of surface-enhanced Raman spectroscopy (SERS) for rapid quantification of the concentration of EAAs in aqueous silver colloids. The EAAs glutamate (Glu) and aspartate (Asp) are released following head injury and have been observed to exhibit SERS spectra that should enable them to be distinguished in a complex aqueous media. Of the two EAAs, the concentration of Glu has been shown to be more indicative of injury to the central nervous system. Using 30-s scans and a 50-mW argon laser, aqueous Glu is quantifiable from 0.4 to 5 micromol/L and is spectrally distinguishable from Asp. In addition, initial in vivo microdialysis experiments suggest that this SERS system is capable of measuring chemical changes following head trauma in the rat brain. Compared with current high-performance liquid chromatography (HPLC) techniques for amino acid detection, the short scanning and processing time associated with the SERS approach enables measurement on a near-real-time basis, providing clinical information in anticipation of pharmaceutical intervention.

Animals↗

PubMed2002

Effect of temperature, pH, and corneal birefringence on polarimetric glucose monitoring in the eye.

Over the last two decades polarimetry has been investigated as a noninvasive alternative for glucose monitoring in support of diabetic patients. In particular, the anterior chamber of the eye containing the fluid known as the aqueous humor has been confirmed to be the optimal sensing site for polarimetric glucose measurements due to its reasonable pathlength (1 cm), low scatter, and minimal depolarization index. In essence, the eye can be thought of as an optical window into the body. In this paper, we will first introduce the key challenges that must be overcome to make the use of polarized light in the eye a viable method for noninvasive glucose monitoring, summarize our work toward this endeavor, and then report on our latest research, namely, the effect of temperature, pH, and corneal birefringence on our polarimetric glucose monitoring system.

Animals↗

PubMed2002

Development and calibration of an automated Mueller matrix polarization imaging system.

The high fatality rate associated with the late detection of skin cancer makes early detection crucial in preventing death. The current method for determining if a skin lesion is suspect to cancer is initially based on the patient's and physician's subjective observation of the skin lesion. Physicians use a set of parameters called the ABCD (asymmetry, border, color, diameter) rule to help facilitate diagnosis of potential cancerous lesions. Lesions that are suspicious then require a biopsy, which is a painful, invasive, and a time-consuming procedure. In an attempt to reduce the aforementioned undesirable elements currently associated with skin cancer diagnosis, a novel optical polarization-imaging system is described that has the potential to noninvasively detect cancerous lesions. The described system generates the full 16-element Mueller matrix in less than 70 s. The operation of the system was tested in transmission, specular reflection, and diffuse reflectance modes, using known samples, such as a horizontal linear polarizer, a mirror, and a diffuser plate. In addition, it was also used to image a benign lesion on a human subject. The results of the known samples are in good agreement with their theoretical values with an average accuracy of 97.96% and a standard deviation of 0.0084, using 16 polarization images. The system accuracy was further increased to 99.44% with a standard deviation of 0.005, when 36 images were used to generate the Mueller matrix.

Biophysical Phenomena↗

PubMedDate not supplied

Detection of differences in oligonucleotide-influenced aggregation of colloidal gold nanoparticles using absorption spectroscopy.

A rapid, simple, and reproducible assay is described that can be used to detect differences in the ability of oligonucleotides to influence the aggregation of colloidal gold nanoparticles. The aggregation reaction of the gold colloid was monitored through UV-visible absorption spectroscopy. Single isolated colloidal gold particles have a surface plasmon resonance manifested as a single absorbance peak at approximately 520 nm, and aggregated gold complexes develop new red-shifted peaks/shoulders depending on the nature and extent of the aggregated complex. A simple ratiometric study of the area under the single and aggregated plasmon resonance peaks thus gives information about the extent of the aggregation. It is postulated that differences in dynamic flexibility of the oligonucleotides affect their influence on the aggregation state of the gold nanoparticles. The results of this study provide new clues toward unraveling the causes behind the preferential affinity of the Hermes transposable element for certain insertion sites compared to other sequences that also contain recognizable target sites. The technique is robust and thus can potentially be used to study similar questions for numerous transposable elements and target sequences.

DNA Mutational Analysis↗

PubMedDate not supplied

Dual-wavelength polarimetry for monitoring glucose in the presence of varying birefringence.

In a continuing effort to develop a noninvasive means of monitoring glucose levels using the aqueous humor of the eye, a dual-wavelength system is developed to show that varying birefringence, similar to what is seen with a moving cornea, can be compensated. In this work, a dual-wavelength, closed-loop system is designed and a model is developed to extract the glucose concentration information. The system and model are tested using various concentrations of glucose in a birefringent test cell subject to motion artifact. The results show that for a static, nonmoving sample, glucose can be predicted to within 10 mg/dl for the entire physiologic range (0 to 600 mg/dl) for either laser wavelength (523 or 635 nm). In the presence of moving birefringence, each individual wavelength produces standard errors on the order of a few thousand mg/dL. However, when the two wavelengths are combined into the developed model, this error is less than 20 mg/dL. The approach shows that multiple wavelengths can be used to drastically reduce the error in the presence of a moving birefringent sample and thus may have the potential to be used to noninvasively monitor glucose levels in vivo in the presence of moving corneal birefringence.

Artifacts↗

PubMedDate not supplied

Measuring microlymphatic flow using fast video microscopy.

Despite advances in the measurement of lymphatic function, little is known about the actual velocities of flow in microlymphatic ( approximately 100 mum diam) vessels. In this work, video microscopy and particle tracking methods are adapted and integrated with an ultra-high-speed imaging camera to obtain measurements of lymph velocities throughout the entire lymphatic contraction cycle in the ratmesentery, something that previous systems were incapable of measuring. To determine the system's accuracy, calibration experiments are conducted across the hypothesized physiologically significant range of velocities for microlymphatic flow (up to 15 mmsec). The system shows high accuracy, less than 2% error, when comparing actual with measured velocities. Microspheres flowing through 140-mum-diam tubing are imaged to demonstrate the system's ability to determine flow rates in these small vessels by measuring particle velocities. To demonstrate biological applicability, mesenteric microlymphatics in loops of the small intestine of three male Sprague-Dawley rats are exteriorized and imaged with the high-speed system at a rate of 500 framessec for several contraction sequences. Lymph velocity fluctuates cyclically with the vessel wall contractions, ranging from -1 to 7 mmsec. These rates are higher than would be possible with standard video microscopy (3.75 mmsec maximum).

Animals↗

PubMedDate not supplied

Use of surface-enhanced Raman spectroscopy for the detection of human integrins.

Current research has revealed the importance of a class of cell surface proteins called integrins in various vital physiological functions such as blood clotting, regulation of blood pressure, tissue blood flow, and vascular remodeling. The key to integrin functionality is its ability to mediate force transmission by interacting with the extracellular matrix and cytoskeleton. In addition, they play a role in signal transduction via their connection with the proteins in focal adhesion (FA) points. To understand the complex mechanism of cell-cell and cell-extracellular matrix (ECM) adhesion that is responsible for these diverse biochemical interactions, it is necessary to identify the integrins on cells and monitor their interaction with various ligands. To this end, for the first time, we employ surface-enhanced Raman spectroscopy (SERS) to detect integrins. The results show the capability using SERS to detect the integrins to the nanomolar concentration regime and to distinguish between two different kinds of integrins, alphaVbeta3 and alpha5beta1, that are present in vascular smooth muscle cells (VSMCs). It is anticipated that the SERS approach will potentially help elucidate the mechanism of integrin-ligand interactions in a variety of phenomena of physiological importance.

Colloids↗