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Biomedical subjects

D J Bornhop

Publications and source records attributed to D J Bornhop.

10 recordsLinked to original sources

Noninvasive picoliter volume thermometry based on backscatter interferometry.

Using the on-chip refractive index (RI) detector based on backscatter interferometry, sensitive, small volume, noninvasive thermometry can be performed. The current optical configuration for the on-chip interferometric backscatter detector (OCIBD) is quite simple and consists of an unfocused laser, an unaltered chip with a hemispherical channel and a photodetector. Alignment is straightforward with the only requirement being that the beam fully fills the channel. The interaction of an unfocused laser beam with the uncoated etched channel with a curvature within the silica plate (chip) produces fringes whose positional changes scale with respect to the refractive index (RI), n, of the fluid in the channel. Due to the inherently high value of dn/dT for most fluids and the high sensitivity of OCIBD to RI changes, the measurement of small temperature variations in sub-nanoliter volumes is possible. Performing OCIBD with a 75 microm diameter laser beam on a silica chip that contains an etched channel with a 40 microm radius facilitates noninvasive thermometry on a N-(2-hydroxyethyl)piperazine-(2-ethanesulfonic acid) (HEPES) solution in a 188 x 10(-12) L probe volume with a temperature resolution of 9.9 x 10(-4) degrees C, at the 99% confidence level.

HEPES↗

Nanoliter-scale non-invasive flow-rate quantification using micro-interferometric back-scatter and phase detection.

The current trend toward miniaturization of fluid-handling systems, particularly those of micro-fluidic devices on the capillary-scale, will certainly lead to improvements in chemical and biochemical analyses. Unfortunately, when fluid volumes reach nano- and picoliter scale it is problematic to perform non-invasive fast and accurate volume flow or flow velocity measurements. Here a simple, non-invasive method is presented for detecting and measuring linear flow velocity within fluid-filled capillaries. A small fluid volume is repeatedly heated locally by means of an infrared laser diode and using the micro-interferometric back-scatter detector (MIBD) at a fixed distance downstream, a thermally induced change in refractive index is observed when the heated volume traverses the probe volume of the detector. Fluid velocity is calculated by monitoring the phase difference between the second harmonic of the heating function and the resulting MIBD output in the Fourier domain. In a probe volume of 40 nL flow rates between I and 10 microL min(-1) are quantifiable, with 3sigma detection limits determined to be 42.8 nL min(-1).

Journal Article↗

Capillary-scale polarimetry for flowing streams.

A micro-polarimeter with a 40 nL probe volume was configured so that it is compatible with capillary-scale flowing stream analysis. The optical configuration consists of two polarizing optics, a capillary, a laser source and a photodetector which is very simple to configure with low cost components. This unique polarimeter is based upon the interaction of a linearly polarized laser beam and a capillary tube, in this case one with an inner diameter of 250 microns. Side illumination of the tube results in a 360 degrees fan of scattered light, which contains a set of interference fringes that change in response to optically active solutes. Solutes that exhibit optical activity are quantifiable and are detected by analyzing the polarization state of the backscattered light. The ability of the instrument to make extremely sensitive optical activity measurements in flowing streams is shown by the determination of (R)-mandelic acid, with a detection limit of 66 x 10(-6) M (507 x 10(-12) g), and the non-optically active control, glycerol. Additionally, the detector was configured to minimize refractive index perturbations.

Microchemistry↗

Detection in capillary electrophoresis.

A review of the four major, on-line, capillary electrophoresis (CE) detection modalities is presented. It is shown that each detection method, fluorescence, absorbance (conventional and nonconventional), electrochemical and refractive index, have distinct advantages and limitations when applied to analysis in a CE format. Various aspects of CE detection are considered and a perspective regarding the applicability of the technique is provided. It is shown that because of widely varying detection limits (ranging from single molecule to 10(-5) M) and detection scheme complexity, the particular application should dictate the selection of detection methodology in CE.

Electrochemistry↗

A chip-scale universal detector for electrophoresis based on backscattering interferometry.

An on-chip detector based on backscatter interferometry has been developed to perform sub-nanoliter volume refractive index measurements. The detection system consists of a simple, folded optical train based on the interaction of a laser beam and an etched channel in a silica (glass) plate. This etched channel is composed of two radii joined by a flat portion which define a curved surface in the shape of a half cylinder in a silica (glass) plate. The backscattered light from the channel takes on the form of a high contrast interference pattern that contains information related to the bulk properties of the fluid located within the probe volume. Positional changes of the interference pattern extrema (fringes) allow for the determination of refractive index changes at the 10(-6) level in a detection volume of 188 x 10(-12) L. Under capillary electrophoresis (CE) conditions, the injected mass detection limits for small molecules with little native absorption ranges from 530 fmol (0.18 ng) for sucrose to 720 fmol (0.43 nanograms) for raffinose. Fluorescein was also used to evaluate the technique for universal CE and under further optimized conditions can be quantified at the 150 microM level. Separation performance for the solutes tested ranged from about 2300 to 15,500 plates or 61,000 to 400,000 N m-1. The results presented here indicate there is potential for using the simple optical train of backscattering interferometry for on-chip universal solute analysis.

Electrophoresis, Capillary↗

Fluorescent tissue site-selective lanthanide chelate, Tb-PCTMB for enhanced imaging of cancer.

In-vivo and in-vitro investigations indicate that a newly developed polyazamacrocyclic chelate of Tb(III) has superior properties for use as an abnormal tissue marker. In addition to tissue selectivity, this molecule is unique because of its low toxicity, attractive fluorescent properties, rapid pharmokinetics, and relatively high water solubility. The complex Tb-3,6,9-tris(methylene phosphonic acid n-butyl ester)-3,6,9,15-tetraazabicyclo[9.3.1]-pentadeca-1(15),11,13 -triene (Tb-PCTMB) has also been shown to exhibit strongly shifted emission (delta lambda--280 nm), moving the detection frequency away from autofluorescence backgrounds, and good quantum efficiencies (phi = 0.51), providing high brightness. Fluorescence imaging was used to quantify Tb-PCTMB at the picomolar level in tissues and to show the significant difference in affinity for the chelate by adenocarcinoma cells HT-29 versus normal epithelial cells (IEC-6). Topical application, or lavage introduction, under endoscopy was used to instill a millimolar aqueous solution of Tb-PCTMB into a dimethylhydrizene-treated Sprague Dawley rat large intestine containing a suspect growth. Subsequent in vitro fluorescence detection and standard histological evaluation confirmed enhanced uptake by adenocarcinoma tissue. Semiquantitative signal interrogation was employed to show the potential for using Tb-PCTMB as a contrast enhancement marker for disease detection.

Animals↗

Refractive-index detection by interferometric backscatter in packed-capillary high-performance liquid chromatography.

A simple, inexpensive, micro-volume refractive index detector has been applied to packed capillary high-performance liquid chromatography. Nanoliter detector volumes produced by a unique optical train, based on interferometric backscatter, allows for universal solute detection at the picogram level. System utility is demonstrated by reversed-phase separation of a test mixture containing ppm levels of NaCl, phenol, ethyl benzene and toluene.

Chromatography, High Pressure Liquid↗

Laser-based capillary polarimeter.

A laser-based capillary polarimeter has been configured to allow for the detection of optically active molecules in capillary tubes with a characteristic inner diameter of 250 microm and a 39-nL (10(-9)) sample volume. The simple optical configuration consists of a HeNe laser, polarizing optic, fused-silica capillary, and charge-coupled device (CCD) camera in communication with a laser beam analyzer. The capillary scale polarimeter is based on the interaction between a polarized laser beam and a capillary tube, which results in a 360 degree fan of scattered light. This array of scattered light contains a set of interference fringe, which respond in a reproducible manner to changes in solute optical activity. The polarimetric utility of the instrument will be demonstrated by the analysis of two optically active solutes, R-mandelic acid and D-glucose, in addition to the nonoptically active control, glycerol. The polarimetric response of the system is quantifiable with detection limits facilitating 1.7 x 10(-3) M or 68 x 10(-12) nmol (7 psi 10(-9) g) sensitivity.

Calibration↗