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Biomedical subjects

Tuan Vo-Dinh

Publications and source records attributed to Tuan Vo-Dinh.

At least 19 recordsLinked to original sources

Time-resolved optical tomography using short-pulse laser for tumor detection.

Our objective is to perform a comprehensive experimental and numerical analysis of the short-pulse laser interaction with a tissue medium with the goal of tumor-cancer diagnostics. For a short-pulse laser source, the shape of the output signal is a function of the optical properties of the medium, and hence the scattered temporal optical signal helps in understanding the medium characteristics. Initially experiments are performed on tissue phantoms embedded with inhomogeneities to optimize the time-resolved optical detection scheme. Both the temporal and the spatial profiles of the scattered reflected and transmitted optical signals are compared with the numerical modeling results obtained by solving the transient radiative transport equation using the discrete ordinates technique. Next experiments are performed on in vitro rat tissue samples to characterize the interaction of light with skin layers and to validate the time-varying optical signatures with the numerical model. The numerical modeling results and the experimental measurements are in excellent agreement for the different parameters studied. The final step is to perform in vivo imaging of anesthetized rats with tumor-promoting agents injected inside skin tissues and of an anesthetized mouse with mammary tumors to demonstrate the feasibility of the technique for detecting tumors in an animal model.

Animals↗

Hyperspectral imaging system using acousto-optic tunable filter for flow cytometry applications.

A major advantage of flow cytometry is its flexible and open instrument configuration, which is highly suitable for systems integration. This flexibility permits the coupling of auxiliary instrumentation that may offer the measurement of parameters other than those typically measured by this multiparameter measurement technique. On the basis of this advantage, we explore the principle and application of hyperspectral imaging (HSI), which has the potential to be a useful add-on feature to flow cytometry applications. Application of HSI to flow cytometry involves the acquisition of spatial information and rendering it in spectral form. In this work, we describe the development and application of an HSI system which provides both spectral and spatial information. Spectral information was generated by obtaining an entire spectrum of a single sample site within a wavelength region of interest, while spatial information was generated by recording a two-dimensional (2D) image of an area of the sample of interest at one specific wavelength. HSI is a promising additional feature to flow cytometry since it can provide both spatial (image format) and spectral information in addition to the multiparameter information already available from flow cytometry measurements.

Acoustics↗

Self-assembly of silver nanoparticles: synthesis, stabilization, optical properties, and application in surface-enhanced Raman scattering.

Silver nanoparticle aggregates were synthesized in large scale using resorcinol under alkaline condition to obtain an assembly of silver clusters. Stable dispersion of the cluster in aqueous medium has been examined out of resorcinol-capped silver nanoparticle assemblies. The UV-vis spectroscopy during the particle evolution has been studied in detail. From the high-resolution TEM (HRTEM) image and XRD pattern it was confirmed that the particles are made of pure silver only. The capping action of resorcinol has been authenticated from the FTIR spectra. UV-vis spectroscopy and TEM images reveal that the temperature, effect of vibrational energy, heat shock, and time-dependent particle evolution have unique bearing on the stability and surface properties of the clusters. The concentrations of silver nitrate, resorcinol, and NaOH have important influence on the particle evolution and its size. TEM images incite us to examine the aggregates to capitulate surface-enhanced Raman scattering (SERS) to the single molecular level using crystal violet (CV) and cresyl fast violet (CFV) as molecular probes. The SERS intensity of CV increases with increasing the size of the silver aggregate.

Metal Nanoparticles↗

Development of an advanced hyperspectral imaging (HSI) system with applications for cancer detection.

An advanced hyper-spectral imaging (HSI) system has been developed having obvious applications for cancer detection. This HSI system is based on state-of-the-art liquid crystal tunable filter technology coupled to an endoscope. The goal of this unique HSI technology being developed is to obtain spatially resolved images of the slight differences in luminescent properties of malignant versus non-malignant tissues. In this report, the development of the instrument is discussed and the capability of the instrument is demonstrated by observing mouse carcinomas in-vivo. It is shown that the instrument successfully distinguishes between normal and malignant mouse skin. It is hoped that the results of this study will lead to advances in the optical diagnosis of cancer in humans.

Animals↗

Detection of human immunodeficiency virus type 1 DNA sequence using plasmonics nanoprobes.

This paper describes the use of plasmonics-based nanoprobes that act as molecular sentinels for DNA diagnostics. The plasmonics nanoprobe comprises a metal nanoparticle and a stem-loop DNA molecule tagged with a Raman label. The nanoprobe utilizes the specificity and selectivity of the DNA hairpin probe sequence to detect a specific target DNA sequence of interest. In the absence of target DNA, the stem-loop configuration maintains the Raman label in proximity to the metal nanoparticle, inducing an intense surface-enhanced Raman scattering (SERS) effect that produces a strong Raman signal upon laser excitation. Upon hybridization of a complementary target DNA sequence to the nanoprobe, the stem-loop configuration is disrupted, causing the Raman label to physically separate from the metal nanoparticle, thus quenching the SERS signal. The usefulness and potential application of the plasmonics nanoprobe for diagnosis is demonstrated using the gag gene sequence of the human immunodeficiency virus type 1 (HIV-1). We successfully demonstrated the specificity and selectivity of the plasmonics nanoprobes to detect PCR amplicons of the HIV gene. The potential for combining the spectral selectivity and high sensitivity of the SERS process with inherent molecular specificity of DNA hairpins to diagnose molecular target sequences in homogeneous solutions is discussed.

Base Sequence↗

An AOTF-based dual-modality hyperspectral imaging system (DMHSI) capable of simultaneous fluorescence and reflectance imaging.

An acousto-optic tunable filter (AOTF)-based system for dual-modality hyperspectral imaging (DMHSI) has been developed for use in characterization of normal and malignant mouse tissue. The system consists of a laser, endoscope, AOTF, and two cameras coupled with optics and electronics. Initial results show that the system can delineate normal and malignant mouse tissues real-time. The analysis shows that malignant tissues consistently exhibit less fluorescent intensity in the wavelength band from 440 to 540 nm with a peak intensity of around 490 nm. The analysis also shows key spectroscopic differences between normal and malignant tissues. Further, these results are compared to real-time spectroscopic data and show good correlation.

Animals↗

Near-real-time determination of hydrogen peroxide generated from cigarette smoke.

The ability to monitor hydrogen peroxide (H2O2) in aqueous smoke extracts will advance our understanding of the relationship between cigarette smoke-induced oxidative stress, inflammation, and disease and help elucidate the pathways by which the various smoke constituents exert their pathogenic effects. We have demonstrated, for the first time, the measurement of H2O2 production from cigarette smoke without prior separation of the sample. Cigarettes were tested on a commercial smoking machine, such that the whole smoke or gas vapor phase was bubbled through phosphate buffered saline solution at pH 7.4. Aliquots of these solutions were analyzed using an Amplex Red/horseradish peroxidase fluorimetric assay that required only a 2 minute incubation time, facilitating the rapid, facile collection of data. Catalase was used to demonstrate the selectivity and specificity of the assay for H2O2 in the complex smoke matrix. We measured approximately 7-8 microM H2O2 from two reference cigarettes (i.e., 1R4F and 2R4F). We also observed 9x more H2O2 from whole smoke bubbled samples compared to the gas vapor phase, indicating that the major constituent(s) responsible for H2O2 formation reside in the particulate phase of cigarette smoke. Aqueous solutions of hydroquinone and catechol, both of which are particulate phase constituents of cigarette smoke, generated no H2O2 even though they are free radical precursors involved in the production of reactive oxygen species in the smoke matrix.

Air Pollutants↗

Fiber-optic nanosensors for single-cell monitoring.

This article is an overview of the fabrication, operating principles, and applications of fiber-optic nanobiosensors with the capability of in-vivo analysis at the single-cell level. Recently, the cross-disciplinary integration of nanotechnology, biology, and photonics has been revolutionizing important areas in molecular biology, especially diagnostics and therapy at the molecular and cellular level. Fiber-optic nanobiosensors are a unique class of biosensor that enable analytical measurements in individual living cells and the probing of individual chemical species in specific locations within a cell. This article provides a review of the research performed in our laboratory and discusses the usefulness and potential of this nanotechnology-based biosensor system in biological research and its applications to biomonitoring of individual cells.

Animals↗

A compact CMOS biochip immunosensor towards the detection of a single bacteria.

Recent use of biological warfare (BW) agents has led to a growing interest in the rapid and sensitive detection of pathogens. Therefore, the development of field-usable detection devices for sensitive and selective detection of BW agents is an important issue. In this work, we report a portable biochip system based on complementary metal oxide semiconductor (CMOS) technology that has great potential as a device for single-bacteria detection. The possibility of single-bacteria detection is reported using an immunoassay coupled to laser-induced fluorescence (LIF) detection. Bacillus globigii spores, which are a surrogate species for B. anthracis spores, were used as the test sample. Enzymatic amplification following immunocomplex formation allowed remarkably sensitive detection of B. globigii spores, and could preclude a complicated optical and instrumental system usually required for high-sensitive detection. Atomic force microscopy (AFM) was employed to investigate whether B. globigii spores detected in the portable biochip system exist in single-cell or multicellular form. It was found that B. globigii spores mostly exist in multicellular form with a small minority of single-cell form. The results showed that the portable biochip system has great potential as a device for single-particle or possibly even single-organism detection.

Bacillus↗

Optical nanobiosensor for monitoring an apoptotic signaling process in a single living cell following photodynamic therapy.

Optical nanobiosensors have enabled bioanalytical measurements to be undertaken within volumes as small as that of single biological cells. In this work, we use nanobiosensors to monitor a molecular signaling process, i.e., caspase-7 activation, following photodynamic therapy (PDT) induced apoptosis in breast cancer cells (MCF-7). PDT induces the mitochondrial pathway of apoptosis which triggers cytochrome c release, activation of caspases-9, -8 and -7 and cleavage of poly (ADP-ribose) polymerase (Parp) protein. Caspase-7 is an important apoptosis-related cysteine protease involved in the activation cascade of caspases and in the proteolytic cleavage of Parp protein. Caspase-7 was detected and identified intracellularly using optical nanobiosensors. Our results show the detection of caspase-7 in single living MCF-cells which in essence typifies the apoptotic event induced by a PDT drug. This work, in principle, demonstrates the minimally invasive capability of optical nanobiosensors to measure important signaling molecules and events in pathways at the single cell level.

Apoptosis↗

Protein nanotechnology: the new frontier in biosciences.

The combination of nanotechnology and molecular biology has led to a new generation of nanoscale-based devices and methods for probing the cell machinery and elucidating intimate life processes occurring at the molecular level that were heretofore invisible to human inquiry. This chapter provides a brief overview of the field of nanotechnology and its applications to the study, design, and use of protein systems in biology and medicine.

Animals↗

Studying 3D subdomains of proteins at the nanometer scale using fluorescence spectroscopy.

Databases devoted to the crystal structure of proteins have dramatically increased in size during the last two decades. Moreover, X-ray and NMR technology studies have shown that proteins belonging to the same family generally share the same global 3D architecture. These results suggest that the need for experimental determination of protein structure will be reduced to those that are suspected to have sufficiently novel structures. Furthermore, NMR and other techniques have demonstrated that a protein in solution experiences constant random thermal motions that occur over large time scales, ranging from picoseconds to seconds and perhaps hours. Such changes may have important functional consequences, but identifying which changes are functionally relevant remains a difficult task even if this problem has been addressed both with experimental and computational methods. For that specific purpose, there is a need for methods allowing a fast and accurate monitoring of conformation changes (that occur at specific sub-domains of proteins. Fluorescence resonance energy transfer (FRET) is a suitable tool for monitoring conformational changes at the nanoscale level. This chapter describes the various FRET methods that are used for monitoring the 3D sub-domain conformation of proteins in solution, in single living cells and at the single molecular level.

Animals↗

Plasmonics-based nanostructures for surface-enhanced Raman scattering bioanalysis.

Surface-enhanced Raman scattering (SERS) spectroscopy is a plasmonics-based spectroscopic technique that combines modern laser spectroscopy with unique optical properties of metallic nanostructures, resulting in strongly increased Raman signals when molecules are adsorbed on or near nanometer-size structures of special metals such as gold, silver, and transition metals. This chapter provides a synopsis of the development and application of SERS-active metallic nanostructures, especially for the analysis of biologically relevant compounds. Some highlights of this chapter include reports of SERS as an immunoassay readout method, SERS gene nanoprobes, near-field scanning optical microscopy SERS probes, SERS as a tool for single-molecule detection, and SERS nanoprobes for cellular studies.

DNA↗

Optical nanosensors for detecting proteins and biomarkers in individual living cells.

Recently, nanotechnology has been revolutionizing important areas in molecular biology and medicine, especially diagnostics and therapy at the molecular and cellular levels. The combination of nanotechnology, biology, advanced materials, and photonics opens up the possibility of detecting and manipulating atoms and molecules using nanodevices. This capability has the potential for a wide variety of medical uses at the cellular level. One of the most recent technological advances has been in the area of nanosensors. This chapter describes the principle of optical nanosensors, their development, and their applications for in vivo analysis of proteins and biomarkers in individual living cells. Nanosensors were fabricated with optical fibers pulled down to tips with distal ends in nanoscale dimensions. Nanosensors with immobilized bioreceptor probes (e.g., antibodies, enzyme substrate) that are selective to target analyte molecules are also referred to as nanobiosensors. Laser light is launched into the fiber, and the resulting evanescent field at the tip of the fiber is used to excite target molecules bound to the antibody molecules. A photometric detection system is used to detect the optical signal (e.g., fluorescence) originating from the analyte molecules or from the analyte-bioreceptor reaction.

Apoptosis↗

Protein amyloidose misfolding: mechanisms, detection, and pathological implications.

A variety of diseases result because of misfolded protein that deposits in extracellular space in the body. These deposits can be amorphous (disordered) or fibrillar (ordered). Inclusion bodies are an example of amorphous aggregates, and amyloid fibril is an example of fibrillar or ordered aggregates. In this chapter, we discuss a class of diseases caused by fibrillar aggregate deposits or amyloid fibrils called amyloidosis. We also review mechanisms by which different proteins misfold to form amyloid fibrils. Each amyloid fibril formed from a different protein causes a different disease by affecting a different organ in the body. However, the characteristics of different amyloid fibrils, namely structure and morphology, observed by electron microscopy and X-ray fiber diffraction appear to be quite similar in nature. We present therapeutic strategies developed to eliminate amyloid fibril formation. These strategies could possibly avert a whole class of fatal diseases caused by amyloid fibril deposition owing to similar characteristics of the amyloid fibrils.

Alzheimer Disease↗

Near-field scanning optical microscopy for bioanalysis at nanometer resolution.

The nondestructive imaging of biomolecules in nanometer domains in their original location and position as adsorbed or deposited on a surface is of garners considerable experimental interest. Near-field scanning optical microscopy (NSOM) is an emerging technique with its astonishing resolving power of <100-nm domains, and nondestructive nature compared with other scanning probe microscopic techniques is an emerging technique to achieve this goal. At the single-molecule level of resolution, it is possible to use the NSOM as a critical tool for visualization of proteins on surfaces to obtain more fundamental information about their orientation and locality without disturbing their original orientation and position, and level of interaction with the surface. Several areas of science and medicine can benefit from this type of study especially for biomedical and biochip applications. To illustrate possible applications, imaging of green fluorescent proteins and biomolecules associated with multidrug resistance proteins in tumor cells will be demonstrated using NSOM.

Animals↗

Detection of cytochrome C in a single cell using an optical nanobiosensor.

In this work, the intracellular measurement of cytochrome c using an optical nanobiosensor is demonstrated. The nanobiosensor is a unique fiberoptics-based tool which allows the minimally invasive analysis of intracellular components. Cytochrome c is a very important protein to the process which produces cellular energy. In addition, cytochrome c is well-known as the protein involved in apoptosis, or programmed cell death. delta-Aminolevulinic acid (5-ALA) was used to induce apoptosis in MCF-7 human breast carcinoma cells. 5-ALA, a photodynamic therapy (PDT) drug in cells was activated by a HeNe laser beam. After the PDT photoactivation, the release of cytochrome c from the mitochondria to the cytoplasm in a MCF-7 cell was monitored by the optical nanobiosensor inserted inside the single cell and followed by an enzyme-linked immunosorbent assay (ELISA) outside the cell. The combination of the nanobiosensor with the ELISA immunoassay improved the detection sensitivity of the nanobiosensor due to enzymatic amplification. Our results lead to the investigation of an apoptotic pathway at the single cell level.

Aminolevulinic Acid↗

Application of a miniature biochip using the molecular beacon probe in breast cancer gene BRCA1 detection.

We report for the first time the application of a biochip using the molecular beacon (MB) detection scheme. The usability of this biochip novel detection system for the analysis of the breast cancer gene BRCA1 is demonstrated using molecular beacon probes. The MB is designed for the BRCA1 gene and a miniature biochip system is used for detection. The performance of the biochip-MB detection system is evaluated. The optimum conditions for the MB system for highest fluorescence detection sensitivity are investigated for the detection system. The detection of BRCA1 gene is successfully demonstrated in solution and the limit of detection (LOD) is estimated as 70 nM.

Biosensing Techniques↗