Biomedical subjects

Sergio B Mendes

Publications and source records attributed to Sergio B Mendes.

7 recordsLinked to original sources

PubMed2006

Combination of polarized TIRF and ATR spectroscopies for determination of the second and fourth order parameters of molecular orientation in thin films and construction of an orientation distribution based on the maximum entropy method.

This article describes two mathematical formalisms for the determination of the second and fourth order parameters of molecular films using optical spectroscopy. Method A uses polarized total internal reflection fluorescence (TIRF) to calculate the second and fourth order parameters, {P2(cos theta)} and {P4(cos theta)}, using an independently determined value for the angle between the absorption and emission dipoles, gamma. Method B uses {P2(cos theta)} obtained from attenuated total reflectance (ATR) data, along with polarized TIRF measurements to calculate {P4(cos theta)} and {cos2 gamma}. The choice of a specific method should rely on experimental considerations. We also present a method to separate the contributions of substrate surface roughness and dipole orientation with respect to the molecular axis from the spectroscopically determined second and fourth order parameters. Finally, a maximum entropy approach for construction of an orientation distribution from order parameters is compared with the commonly used delta and Gaussian distributions.

Entropy↗

PubMed2006

Order parameters and orientation distributions of solution adsorbed and microcontact printed cytochrome c protein films on glass and ITO.

The structure of solution adsorbed and microcontact printed (muCP) cytochrome c (cyt c) films on glass and indium tin oxide (ITO) was investigated using attenuated total reflectance (ATR) and total internal reflectance fluorescence (TIRF) spectroscopies to determine the orientation of the heme groups in the films. The second and fourth order parameters of the heme as well as information on the angle between the absorption and emission dipoles of the heme, gamma, were experimentally determined. The order parameters of the heme are related to the order parameters of the protein molecule using the known angle between the heme plane and the electrostatic dipole moment of the cyt c protein. The effect of the surface roughness of the substrates (glass and ITO) was also taken into account quantitatively using AFM data. Physically possible order parameters were obtained for the heme group in both solution adsorbed and muCP films, but not for the electrostatic dipole moment of the protein. In addition, the experimental values of {cos2 gamma} for immobilized zinc-substituted cyt c are greater than the values of {cos2 gamma} determined in viscous solutions, which could be an indication that the environment of the heme groups changes upon adsorption. The electron transfer behavior of solution adsorbed and muCP films on ITO, determined using electrochemical methods, is compared to their orientation distribution and surface coverage as determined by spectroscopic methods.

Adsorption↗

PubMed2005

Determination of anisotropic optical constants and surface coverage of molecular films using polarized visible ATR spectroscopy. Application to adsorbed cytochrome c films.

This article describes a method to determine the anisotropic optical constants and surface coverage of molecular films using polarized attenuated total reflectance (ATR) absorbance measurements. We have extended the transfer-matrix formalism to describe birefringent and dichroic films in ATR geometries and have combined it with an iterative numerical procedure to determine the anisotropic values of both the real (n) and imaginary (k) parts of the complex refractive index of the film under investigation. Anisotropic values of the imaginary part of the refractive index (k) allow for the determination of the surface coverage and one order parameter of the film. To illustrate this approach, we have used cytochrome c (cyt c) protein films adsorbed to glass and indium tin oxide (ITO) surfaces. Experimental results show that cyt c films on these surfaces, which were formed under identical conditions, have significant differences in their surface coverages (11.2 +/- 0.4 pmol/cm(2) on glass and 21.7 +/- 0.9 pmol/cm(2) on ITO); however, their order parameters are similar (0.30 +/- 0.02 on glass and 0.36 +/- 0.04 on ITO).

Adsorption↗

PubMed2005

Determination of molecular anisotropy in thin films of discotic assemblies using attenuated total reflectance UV-visible spectroscopy.

We report here an investigation of absorbance anisotropy in highly ordered, single bilayer (ca. 5.6 nm) Langmuir-Blodgett (LB) thin films of discotic liquid-crystalline phthalocyanines, using a recently introduced broad-band attenuated total reflectance (ATR) spectroscopic technique, capable of measuring dichroism in such films in the UV--visible optical region down to absorbances of ca. 0.003 absorbance units. On the basis of the ATR measurements of LB-deposited films, a thorough treatment was established to determine the ensemble average of the Cartesian components and the associated optical anisotropy of transition dipoles in the molecular film. In an effort to recover order parameters of molecular orientation, those results were interpreted with a circular dipole model, which is the expected model for the isolated molecule based on symmetry properties. We measured a strong dipole component normal to the film plane that cannot be explained in terms of a truly circular model, indicating that the molecular transition dipoles were perturbed upon aggregation. The utility of the experimental approach was further demonstrated by (a) investigating the effect of substrate modifiers (methyl- and phenyl-terminated silanes) on the ordering within the phthalocyanine film and (b) the effect of water immersion and re-annealing of the thin film on molecular ordering and optical anisotropy.

Journal Article↗

PubMed2004

Technique for determining the angular orientation of molecules bound to the surface of an arbitrary planar optical waveguide.

A technique to determine the angular orientation of a molecular assembly bound to the surface of a planar optical waveguide of arbitrary structure is described. The approach is based on measuring the absorption dichroic ratio by using the waveguide evanescent fields with orthogonal polarizations (TE, TM) and the same mode order to probe two molecular assemblies, (i) a reference sample composed of an isotropic orientation distribution of dipoles and (ii) a sample of interest. The isotropic sample is used to characterize the waveguide structure, which then allows the orientation parameters of a molecular assembly under investigation to be determined from a measured dichroic ratio. The method developed here is particularly important for applications in gradient-index and multilayer planar waveguide platforms because in those cases the extension of previously reported approaches would require a full experimental characterization of the guiding structure, which would be problematic and may yield inaccurate results.

Journal Article↗

PubMed2003

Broadband coupling into a single-mode, electroactive integrated optical waveguide for spectroelectrochemical analysis of surface-confined redox couples.

A single-mode, electroactive waveguiding platform capable of measuring spectroelectrochemical responses of surface-adsorbed redox-active molecules over a broad spectral bandwidth has been created. This new planar waveguide spectrometer is a combination of the previously developed electroactive integrated optical waveguide (EA-IOW; Dunphy, D. R.; Mendes, S. B.; Saavedra, S. S.; Armstrong, N. R. Anal. Chem. 1997, 69, 3086-3094) with a recently reported simplified approach to broadband coupling (Bradshaw, J. T.; Mendes, S. B.; Saavedra, S. S. Anal. Chem. 2002, 74, 1751-1759). With the use of a commercially available prism as an incoupling element, the EA-IOW can now guide visible light from at least 500 to 700 nm, improving upon its previously demonstrated monochromatic nature. Coupling profiles of various laser lines along with transmission spectra of narrow band-pass filters at various potentials are used to demonstrate the optical characteristics of this broadband EA-IOW and to compare its response to that of a conventional transmission instrument. Assessment of spectral resolution, performed by measuring the fwhm of various laser lines, ranges from 0.6 to 0.8. To demonstrate the capabilities of this technology, we show the acquisition of absorbance spectra of two different adsorbates, cytochrome c and ferrocenedicarboxylic acid, as a function of applied potential. Subtleties in the redox chemistries of adsorbed molecules, which were difficult to monitor with a monochromatic waveguide, are readily apparent when using the broadband coupling scheme.

Electrochemistry↗

PubMed2002

A simplified broadband coupling approach applied to chemically robust sol-gel, planar integrated optical waveguides.

A new generation waveguide spectrometer with broadband coupling capabilities has been developed. As opposed to previous devices, this attenuated total reflection (ATR) spectrometer is much simpler in design, is more chemically robust, and transmits light down to at least 400 nm. The attenuated total reflection element consists of a single-mode, planar integrated optical waveguide fabricated by dip-coating a approximately 300 nm thick, sol-gel composite layer on a glass substrate. A commercially available prism is used as the incoupler with an integral holographic diffraction grating acting as the dispersive outcoupling element. The transmission of narrow band-pass filters was used to compare the response of the waveguide spectrometer to that of a conventional transmission instrument. Spectral resolution was assessed by measuring the fwhm of various laser lines, which were found to range from 0.5 to 1.3 nm. The measured limits of detection for the waveguide spectrometer from 400 to 600 nm are 8.0 and 10.1 milliabsorbance units for TE and TM polarizations, respectively. Finally, to demonstrate the application of this technology to a molecular film confined to a solid-liquid interface, visible ATR spectra of an adsorbed submonolayer of horse heart cytochrome c were acquired. A procedure to correct the waveguide spectra for the wavelength dependence in ATR path length is described.

Animals↗