The SIT image Vidicon as a gas-phase fluorescence detector for gas chromatography.
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Air Pollutants↗
Biomedical subjects
T Vo-Dinh
Publications and source records attributed to T Vo-Dinh.
Fluorescence studies of benzo-[a]-pyrene in liposome membrane systems.
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Benzopyrenes↗
Heavy-atom effect on room temperature phosphorimetry.
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Iodides↗
Development of a new capillary electrophoresis-based fibre optic sensor.
A new fluorescence-based fibre optic sensor is described which combines the sensitivity offered by laser-induced fluorescence with the selectivity offered by capillary electrophoresis (CE). A single optical fibre directly probes the terminus of a 5-8 cm separation capillary. The linear geometry associated with this sensor necessitates a 'single reservoir' design, thus presenting major challenges to overcome in comparison to the conventional two-reservoir configuration common to a typical laboratory setup. Some of the challenges confronted by the design features presented in this work include the reduction of gravity-driven hydrostatic flow, the ejection of electrolytic gases evolved at the detection-side electrode and the establishment a suitable compromise between detectability and separation performance. The success of such design features demonstrates the feasibility of a CE-based sensor which offers several amenities particularly useful for in situ sensing. Such attributes include selectivity, diminutive size, flexibility, reusability, high sensitivity, speed, and remote control. Detailed descriptions of sensor fabrication are included, including two variations on a general design concept. In addition, the single-fibre optical detection system is described. Separation characteristics of the new CE-based sensor are presented, highlighted by an observed separation efficiency of up to 8000 theoretical plates (for a 5 cm capillary). The separation of a three-component mixture of the laser dyes, Rhodamine 6G, fluorescein isothyocyanate and sodium fluorescein, is demonstrated.
Biosensing Techniques↗
Antibody-antigen binding kinetics. A model for multivalency antibodies for large antigen systems.
This work presents a theoretical analysis of the influence of multivalency of antigen on external mass transfer-limited binding kinetics to divalent antibody for biosensor applications to polycyclic-aromatic systems. Both cases are considered wherein the antigen is in solution and the antibody is either covalently or noncovalently attached to a cylindrical fiber-optic biosensor, and the antibody is in solution and the antigen is attached to the surface. Both single-step and dual-step binding processes are considered. The rate of attachment of antigen to antibody (or vice versa) is linear for the valencies (or reaction orders) analyzed in the time frame (100 min) considered. The rate of attainment of saturation levels of antigen or antibody in solution close to the surface is very rapid (within 20 min). An increase in the valency of the antigen in solution has the effect of decreasing the order of reaction (for valency, v > or = 1). An increase in the number of steps increases the order of reaction, as expected. An increase in the valency of the antigen in solution decreases the saturation level of the antigen close to the surface and the rate of antigen attachment to the antibody on the surface for all Damkohler numbers. A decrease in the diffusional limitations decreases the effect of valency (or reaction order) on saturation levels of Cs/C0. Nondimensional plots presented in the analysis help extend the analysis to different antigen-antibody systems. An increase in the valency of the antibody in solution has the effect of increasing the order of reaction (for v > 2). The effects in this case are reverse to those described earlier. For valency greater than 2, the reaction order is dependent on the antigen valency, whether it is in solution or immobilized on the surface. The general analysis presented here should be applicable to most surface reactions that involve ligand-receptor binding wherein multiple-binding sites are involved on either the receptor or the ligand.
Antibodies↗
Biosensors and biochips: advances in biological and medical diagnostics.
In the past two decades, the biological and medical fields have seen great advances in the development of biosensors and biochips capable of characterizing and quantifying biomolecules. This review is meant to provide an overview of the various types of biosensors and biochips that have been developed for biological and medical applications, along with significant advances over the last several years in these technologies. It also attempts to describe various classification schemes that can be used for categorizing the different biosensors and provide relevant examples of these classification schemes from recent literature.
Biosensing Techniques↗
Application of an antibody biochip for p53 detection and cancer diagnosis.
Detection of the p53 tumor suppressor gene is important in early cancer diagnostics because alterations in the gene have been associated with carcinogenic manifestations in several tissue types in humans. We have developed an antibody-based detection instrument, the biochip, to detect the presence of the anti-p53 antibody in human serum. The design of this highly integrated detector system is based on miniaturized phototransistors having multiple optical sensing elements, amplifiers, discriminators, and logic circuitry on an IC board. The system utilizes laser excitation and fluorescence signals to detect complex formation between the p53 monoclonal antibody and the p53 antigen. Recognition antibodies are immobilized on a nylon membrane platform and incubated in solutions containing antigens labeled with Cy5, a fluorescent cyanine dye. Subsequently, this membrane is placed on the detection platform of the biochip and fluorescence signal is induced using a 632.8-nm He-Ne laser. Using this immuno-biochip, we have been able to detect binding of the p53 monoclonal antibody to the human p53 cancer protein in biological matrices. The performance of the integrated phototransistors and amplifier circuits of the biochip, previously evaluated through measurement of the signal output response for various concentrations of fluorescein-labeled molecules, have illustrated the linearity of the microchip necessary for quantitative analysis. The design of this biochip permits sensitive, selective and direct measurements of a variety of antigen-antibody formations at very low concentrations. Furthermore, the acquisitions of the qualitative and quantitative results are accomplished rapidly, in about 15 min. These features demonstrate the potential of this antibody-based biochip for simple, rapid and early biomedical diagnostics of cancer.
Antibodies, Monoclonal↗
Single- and dual-fractal analysis of hybridization binding kinetics: biosensor applications.
The diffusion-limited hybridization kinetics of analyte in solution to a receptor immobilized on a biosensor or immunosensor surface is analyzed within a fractal framework. The data may be analyzed by a single- or a dual-fractal analysis. This was indicated by the regression analysis provided by Sigmaplot. It is of interest to note that the binding rate coefficient and the fractal dimension both exhibit changes in the same direction for both the single-fractal and the dual-fractal analysis examples presented. For example, for a single-fractal analysis and for the hybridization of 10 nM 16CFl (oligonucleotide) to 16B immobilized via sulfosuccinimidyl-6-(biotinamido)hexanoate and streptavidin using chemical and thermal regeneration (Abel, A. P.; Weller, M. G.; Duveneck, G. L.; Ehrat, M. Widmer, H. M. Anal. Chem. 1996, 68, 2905-2912), an increase in the fractal dimension, Df from 1.211 (chemical regeneration) to 1.394 (thermal regeneration), leads to an increase in the binding rate coefficient, k, from 86.53 (chemical regeneration) to 100.0 (thermal regeneration). An increase in the degree of heterogeneity on the biosensor surface leads to an increase in the binding rate coefficient. When a dual-fractal analysis was utilized, an increase in the fractal dimension value from Df1 to Df2 leads to an increase in the binding rate coefficient value from k1 to k2. The fractional order of dependence of the binding rate coefficient, k1, on (a) the analyte (rRNA) concentration in solution and (b) on the fractal dimension, Df1, for the hybridization kinetics to detect Listeria species (Fliss, R.; St-Laurent, M.; Emond, E.; Simard, R. E.; Lemieux, R.; Ettriki, A.; Pandian, S. Appl. Microbiol. Biotechnol. 1995, 43, 717-724.) further reinforces the fractal nature of the system. The binding rate coefficient(s) expressions developed as a function of the analyte concentration in solution and the fractal dimension are of particular value since they provide a means to better control of biosensor or immunosensor performance.
Biosensing Techniques↗
Measurement of DNA adducts using surface-enhanced Raman spectroscopy.
Hazardous pollutants emitted from energy-related technologies, chemical industries, or waste materials are of increasing public concern because of their potential adverse health effects. Many pollutants have chemical groups of toxicological importance that can be characterized and detected by Raman spectroscopy. Raman spectroscopy, however, has not been widely used in trace organic detection, even though the information contained in a Raman spectrum is valuable for chemical identification. One limitation of conventional Raman spectroscopy is its low sensitivity, which often necessitates the use of powerful and costly laser sources for sample excitation. Raman spectroscopists have recently been able to analyze dilute biological samples as a result of enhancements in the Raman scattering cross section by factors up to 10(10) when a compound is adsorbed on or near a special electron-conducting surface. These spectacular enhancement factors of the normally weak Raman scattering process help overcome the low sensitivity of Raman spectroscopy through a combination of electromagnetic and chemical interactions between the analyte molecule and the surface. The technique associated with this phenomenon is known as surface-enhanced Raman scattering spectroscopy (SERS). The special conductive surface responsible for the scattering enhancement is referred to as a SERS substrate. For the past few years we have developed the SERS technique, using practical SERS-active substrate materials based on silver-coated microspheres deposited on glass. A wide variety of biomarkers have been investigated, including benzo[a]pyrene, dibenz[a,h]anthracene epoxides, 1,N9-ethenoadenine, 3,N4-ethenocytosine, and other substances. These biomarkers were measured at nanogram and subnanogram levels. The experimental results are of great analytical interest, since these chemicals are difficult to detect by other techniques, such as luminescence spectroscopy, because of the weak luminescence quantum yields of these DNA adducts. In this paper the potential usefulness of the SERS technique for assessing environmental and health effects from human exposure to toxic pollutants is demonstrated.
Animals↗
Discrimination between tumour and normal cells by staining with 3,4,5,6,16,17-hexadehydro-16-(methoxycarbonyl)-19 alpha-methyl-20 alpha-oxayohimbanium: the uracil ring as a target for the specific interaction between RNA(s) and the fluorescent probe.
3,4,5,6,16,17-Hexadehydro-16-(methoxycarbonyl)-19 alpha-methyl-20 alpha-oxayohimbanium (Alstonine) is a fluorescent alcaloid which has been known to stain tumour cells more efficiently than normal ones. In this paper the spectral properties of Alstonine were first investigated and its capability for preferential staining of tumour cells verified in culture using SK-OV-3 cells as tumour cells and Mouse 3T3 fibroblasts as controls. Then interactions between Alstonine and biological macromolecules were investigated to provide the rationale for preferential labelling. Molecular filtration techniques have demonstrated that binding occurs only with RNA molecules. Similar experiments were performed with different isopolynucleotides to find an explanation for that specificity. They provide evidence that binding occurs only in the presence of a uridyl ring. This is consistent with the specificity of the linkage to RNA. As the linkage of Alstonine with RNA did not induce any shift or obvious change in the intensity of its fluorescence spectrum, it is concluded that the binding might involve the side chain of the fluorescent compound.
3T3 Cells↗