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Biomedical subjects

Gordon M Miskelly

Publications and source records attributed to Gordon M Miskelly.

6 recordsLinked to original sources

Reflective interferometric fourier transform spectroscopy: a self-compensating label-free immunosensor using double-layers of porous SiO2.

An interferometric biosensor comprised of two layers of porous Si, stacked one on top of the other, is described. A fast Fourier transform (FFT) of the reflectivity spectrum reveals three peaks that correspond to the optical thickness of the top layer, the bottom layer, and both layers together. Binding of immunoglobulin G to a protein A capture probe adsorbed to the surface of the top layer induces changes in reflectivity at the top layer/solution interface. The FFT method allows discrimination of target analyte binding from matrix effects due to nonspecific changes in the analyte solution. The sensor response is shown to be insensitive to the addition of 4000-fold excess sucrose or 80-fold excess bovine serum albumin interferents.

Animals↗

Biosensing using porous silicon double-layer interferometers: reflective interferometric Fourier transform spectroscopy.

A simple, chip-based implementation of a double-beam interferometer that can separate biomolecules based on size and that can compensate for changes in matrix composition is introduced. The interferometric biosensor uses a double-layer of porous Si comprised of a top layer with large pores and a bottom layer with smaller pores. The structure is shown to provide an on-chip reference channel analogous to a double-beam spectrometer, but where the reference and sample compartments are stacked one on top of the other. The reflectivity spectrum of this structure displays a complicated interference pattern whose individual components can be resolved by fitting of the reflectivity data to a simple interference model or by fast Fourier transform (FFT). Shifts of the FFT peaks indicate biomolecule penetration into the different layers. The small molecule, sucrose, penetrates into both porous Si layers, whereas the large protein, bovine serum albumin (BSA), only enters the large pores. BSA can be detected even in a large (100-fold by mass) excess of sucrose from the FFT spectrum. Detection can be accomplished either by computing the weighted difference in the frequencies of two peaks or by computing the ratio of the intensities of two peaks in the FFT spectrum.

Biosensing Techniques↗

Using spectral information in forensic imaging.

Improved detection of forensic evidence by combining narrow band photographic images taken at a range of wavelengths is dependent on the substance of interest having a significantly different spectrum from the underlying substrate. While some natural substances such as blood have distinctive spectral features which are readily distinguished from common colorants, this is not true for visualization agents commonly used in forensic science. We now show that it is possible to select reagents with narrow spectral features that lead to increased visibility using digital cameras and computer image enhancement programs even if their coloration is much less intense to the unaided eye than traditional reagents. The concept is illustrated by visualising latent fingermarks on paper with the zinc complex of Ruhemann's Purple, cyanoacrylate-fumed fingerprints with Eu(tta)(3)(phen), and soil prints with 2,6-bis(benzimidazol-2-yl)-4-[4'-(dimethylamino)phenyl]pyridine [BBIDMAPP]. In each case background correction is performed at one or two wavelengths bracketing the narrow absorption or emission band of these compounds. However, compounds with sharp spectral features would also lead to improved detection using more advanced algorithms such as principal component analysis.

Chlorides↗

Inhibition of bleach-induced luminol chemiluminescence.

The luminol chemiluminescence presumptive test for blood is based on the mild peroxidase activity of hemoglobin in basic peroxide solution. However, this test is subject to interference by strong oxidants, certain transition metal ions, and true peroxidases. This paper reports methods for reducing the interference caused by hypochlorite-containing bleaches. Amines such as 1,2-diaminoethane react rapidly with hypochlorite without interfering significantly with the hemoglobin-catalyzed oxidation. Thus, addition of 0.1 mol/L 1,2-diaminoethane to a standard luminol-peroxide spray lead to almost complete inhibition of hypochlorite-induced chemiluminescence while satisfactory chemiluminescence was still observed from bloodstains. If time allows, an alternative method for reducing interference from hypochlorite bleach is to wait several days until the bloodstains have dried thoroughly, by which time the hypochlorite will have decomposed.

Blood Stains↗

Background correction in forensic photography. I. Photography of blood under conditions of non-uniform illumination or variable substrate color--theoretical aspects and proof of concept.

The combination of photographs taken at two or three wavelengths at and bracketing an absorbance peak indicative of a particular compound can lead to an image with enhanced visualization of the compound. This procedure works best for compounds with absorbance bands that are narrow compared with "average" chromophores. If necessary, the photographs can be taken with different exposure times to ensure that sufficient light from the substrate is detected at all three wavelengths. The combination of images is readily performed if the images are obtained with a digital camera and are then processed using an image processing program. Best results are obtained if linear images at the peak maximum, at a slightly shorter wavelength, and at a slightly longer wavelength are used. However, acceptable results can also be obtained under many conditions if non-linear photographs are used or if only two wavelengths (one of which is at the peak maximum) are combined. These latter conditions are more achievable by many "mid-range" digital cameras. Wavelength selection can either be by controlling the illumination (e.g., by using an alternate light source) or by use of narrow bandpass filters. The technique is illustrated using blood as the target analyte, using bands of light centered at 395, 415, and 435 nm. The extension of the method to detection of blood by fluorescence quenching is also described.

Biophysical Phenomena↗

Background correction in forensic photography. II. Photography of blood under conditions of non-uniform illumination or variable substrate color--practical aspects and limitations.

The combination of photographs taken at wavelengths at and bracketing the peak of a narrow absorbance band can lead to enhanced visualization of the substance causing the narrow absorbance band. This concept can be used to detect putative bloodstains by division of a linear photographic image taken at or near 415 nm with an image obtained by averaging linear photographs taken at or near 395 and 435 nm. Nonlinear images can also be background corrected by substituting subtraction for the division. This paper details experimental applications and limitations of this technique, including wavelength selection of the illuminant and at the camera. Characterization of a digital camera to be used in such a study is also detailed. Detection limits for blood using the three wavelength correction method under optimum conditions have been determined to be as low as 1 in 900 dilution, although on strongly patterned substrates blood diluted more than twenty-fold is difficult to detect. Use of only the 435 nm photograph to estimate the background in the 415 nm image lead to a twofold improvement in detection limit on unpatterned substrates compared with the three wavelength method with the particular camera and lighting system used, but it gave poorer background correction on patterned substrates.

Biophysical Phenomena↗