Hadamard transform microchip electrophoresis combined with diode laser fluorometry

The application of a Hadamard transform technique to microchip electrophoresis is described. The sample is electrokinetically injected into a separation channel and is then detected by diode laser-induced fluorometry. The sample and buffer solutions are introduced into the channel by controlling the high voltages applied to the solutions, according to a code determined by a Hadamard matrix. The S/N ratio of the signal in the electropherogram can be improved by a factor of 5 in comparison with that obtained by a conventional single-injection method, although an 8-fold improvement is theoretically predicted when a 255-order matrix is used.     

Hadamard transform capillary electrophoresis combined with absorption spectrometry

A novel injection device for applying absorption spectrometry to Hadamard transform (HT) capillary electrophoresis is described. A small hole, at the center part of the capillary, functions as an inlet port for the sample. The hole is immersed in a sample solution and the end of the capillary that is usually employed for sample introduction is immersed in a buffer solution. An ultraviolet absorption detector is placed between the sample injection port and the other end of the capillary filled with a buffer solution. A high potential is continuously applied between the injection port and the end of the capillary, which allows the sample solution to be introduced into the separation capillary. By application of a higher potential modulated according to a Hadamard code between both ends of the capillary, the buffer solution is injected into the separation capillary. In some preliminary experiments, this injection device was utilized to introduce a single sample segment into a capillary. As expected, a single peak was observed in the electropherogram for a sample containing a single component. This device was then employed for multiple sample injection in HT capillary electrophoresis. An 8-fold improvement in the S/N ratio was observed when the HT technique was used, in which a 255-order of a Hadamard matrix was used, as expected from theory. The present approach was also utilized for the sensitive detection of a sample comprised of multiple components.     

Dual-electrode electrochemical detection for poly(dimethylsiloxane)-fabricated capillary electrophoresis microchips

The development of a poly(dimethylsiloxane)-based (PDMS-based) microchip electrophoresis system employing dual-electrode electrochemical detection is described. This is the first report of dual-electrode electrochemical detection in a microchip format and of electrochemical detection on chips fabricated from PDMS. The device described in this paper consists of a top layer of PDMS containing the separation and injection channels and a bottom glass layer onto which gold detection electrodes have been deposited. The two layers form a tight reversible seal, eliminating the need for high-temperature bonding, which can be detrimental to electrode stability. The channels can also be temporarily removed for cleaning, significantly extending the lifetime of the chip. The performance of the chip was evaluated using catechol as a test compound. The response was linear from 10 to 500 microM with an LOD (S/N = 3) of 4 microM and a sensitivity of 45.9 pA/microM. Collection efficiencies for catechol ranged from 28.7 to 25.9% at field strengths between 200 and 400 V/cm. Dual-electrode detection in the series configuration was shown to be useful for the selective monitoring of species undergoing chemically reversible redox reactions and for peak identification in the electropherogram of an unresolved mixture.     

Microchip capillary electrophoresis with an integrated indium tin oxide electrode-based electrochemiluminescence detector

This paper describes an indium tin oxide (ITO) electrode-based Ru(bpy)3(2+) electrochemiluminecence (ECL) detector for a microchip capillary electrophoresis (CE). The microchip CE-ECL system described in this article consists of a poly(dimethylsiloxane) (PDMS) layer containing separation and injection channels and an electrode plate with an ITO electrode fabricated by a photolithographic method. The PDMS layer was reversibly bound to the ITO electrode plate, which greatly simplified the alignment of the separation channel with the working electrode and enhanced the photon-capturing efficiency. In our study, the high separation electric field had no significant influence on the ECL detector, and decouplers for isolating the separation electric field were not needed in the microchip CE-ECL system. The ITO electrodes employed in the experiments displayed good durability and stability in the analytical procedures. Proline was selected to perform the microchip device with a limit of detection of 1.2 microM (S/N = 3) and a linear range from 5 to 600 microM.     

Accurate pixel-to-pixel correspondence adjustment in a digital micromirror device camera by using the phase-shifting moiré method

A camera based on the digital micromirror device (DMD) technology has been previously developed. In this optical system, the correspondence of each mirror of the DMD to each pixel of the CCD cannot readily be done since the pixel sizes of the DMD and the CCD are very small. An accurate pixel-to-pixel correspondence adjustment in the DMD camera by means of the phase-shifting moiré method is proposed. To perform high accurate adjustment of the optical system, the phase distribution of a moiré fringe pattern is analyzed when the CCD pixels and the DMD mirrors have a mismatch and/or misalignment with each other. This technique does not need a complicated setting or complex image processing to generate the moiré fringe pattern, and it needs only one captured image. In the adjustment experiment, the proposed method provided very accurate adjustment whose error was less than 1/25 pixel. An experiment of phase analysis to demonstrate the usefulness was performed.     

Trace analysis of DNA: preconcentration, separation, and electrochemical detection in microchip electrophoresis using Au nanoparticles

We have developed a simple and sensitive on-chip preconcentration, separation, and electrochemical detection (ED) method for trace analysis of DNA. The microchip comprised of three parallel channels: the first two are for the field-amplified sample stacking and subsequent field-amplified sampled injection steps, while the third one is for the microchip gel electrophoresis (MGE) with ED (MGE-ED). To improve preconcentration and separation performances of the method, the stacking and separation buffers containing the hydroxypropyl cellulose (HPC) matrix were modified with gold nanoparticles (AuNPs). The formation of AuNPs and HPC/AuNP-modified buffers were characterized by UV-visible spectroscopy and TEM experiments. The conducting polymer-modified electrode was also modified with AuNPs to enhance detection performances of the electrode. The conducting polymer/AuNP layers act as electrocatalysts for the direct detection of DNA based on their oxidation in a solution phase. The total sensitivity was improved by approximately 25 000-fold when compared with a conventional MGE-ED analysis. The calibration plots were linear (r2 = 0.9993) within the range of 0.003-1.0 pg/microL for a 20-bp DNA sample. The sensitivity was 0.20 nA/(fg/microL), with a detection limit of 5.7 amol in a 50-microL sample, based on S/N = 3. The applicability of the method for the analysis of 13 fragments present in a 100-bp DNA ladder was successfully demonstrated.     

Performance of pyrolyzed photoresist carbon films in a microchip capillary electrophoresis device with sinusoidal voltammetric detection

Pyrolyzed photoresist films (PPF) are introduced as planar carbon electrodes in a PDMS-quartz hybrid microchip device. The utility of PPF in electroanalytical applications is demonstrated by the separation and detection of various neurotransmitters. PPF is found to form a stable, low-capacitance, durable layer on quartz, which can then be used in conjunction with a microchip capillary electrophoretic device. Sinusoidal voltammetric detection at PPF electrodes is shown to be very sensitive, with a detection limit (S/N = 3) of 100 nM for dopamine, corresponding to a mass detection limit (S/N = 3) of 2 amol. The selectivity of analysis in the frequency domain is demonstrated by isolating each individual signal in a pair of analytes that are chromatographically unresolved. Effectively decoupling the electrophoresis and electrochemical systems allows the electrodes to be placed just inside the separation channel, which results in efficient separations (80 000-100 000 plates/m).     

交会跟踪CCD像机探测合作目标光功率分析

以辐射度量为标准,从CCD像机灵敏度出发,建立了CCD像机探测合作目标光功率模型以及功率方程,得到了所需合作目标光功率与CCD像机参数的制约关系,分析了CCD像机探测到光点所占像元数与传输距离之间的关系,为交会跟踪系统的设计和性能评估提供了理论依据。     

Indirect measurement of nitric oxide production by monitoring nitrate and nitrite using microchip electrophoresis with electrochemical detection

An indirect method for monitoring nitric oxide (NO) by determining nitrate and nitrite using microchip capillary electrophoresis (CE) with electrochemical (EC) detection has been developed. This method combines determination of nitrite by direct amperometric detection following a microchip-based CE separation and conversion of nitrate to nitrite by chemical reduction using Cu-coated Cd granules. The amount of nitrate is quantified by calculating the difference in the amount of nitrite in the sample before and after the reduction of nitrate. Optimization of the separation, injection, detection, and reduction reaction conditions, as well as studies involving integration of the reduction reaction onto the microchip, are described. It was found that nitrite can be separated and detected in approximately 45 s by microchip CEEC. The reduction reaction was successfully integrated on-chip and carried out in approximately 1 min following activation of the Cd granules. The usefulness of this device was demonstrated by monitoring the amount of nitrate and nitrite produced from 3-morpholinosydnonimine, a NO-releasing compound.     

Direct analysis of trace phenolics with a microchip: in-channel sample preconcentration, separation, and electrochemical detection

A micrototal analytical method assembling in-channel preconcentration, separation, and electrochemical detection steps has been developed for trace phenolic compounds. A micellar electrokinetic chromatography separation technique was coupled with two preconcentration steps of field-amplified sample stacking (FASS) and field-amplified sample injection (FASI). An amperometric detection method with a cellulose-dsDNA-modified, screen-printed carbon electrode was applied to detect preconcentrated and separated species at the end of the channel. The microchip was composed of three parallel channels: first, two are for the sample preconcentration using FASS and FASI methods, and the third one is for the separation and electrochemical detection. The modification of the electrode surface improved the detection performance by enhancing the signal-to-noise characteristic without surface fouling of the electrode. The method was examined for the analysis of eight phenolic compounds. Experimental parameters affecting the analytical performance of the method were assessed and optimized. The preconcentration factor was increased by about 5200-fold as compared with a simple capillary zone electrophoretic analysis using the same channel. Reproducible response was observed during multiple injections of samples with a RSD of <8.0%. The calibration plots were shown to be linear (with the correlation coefficient between 0.9913 and 0.9982) over the range of 0.4-600 nM. The sensitivity was between 0.17 +/- 0.001 and 0.48 +/- 0.006 nA/nM, with the detection limit of approximately 100 to approximately 150 pM based on S/N = 3. The applicability of the method to the direct analysis of trace phenolic compounds in water samples was successfully demonstrated.     

A microchip electrophoresis device with integrated electrochemical detection: a direct comparison of constant potential amperometry and sinusoidal voltammetry

A microchip electrophoresis system with integrated electrochemical detection is described in this work. The hybrid device utilizes poly(dimethylsiloxane) as the electrophoresis channel substrate and a planar gold electrode lithographically fabricated onto a glass slide for electrochemical detection. The system is characterized by the separation and detection of various neurotransmitters. The gold working electrode is placed just inside the separation channel without adverse effects on the detection sensitivity, due to the electrical decoupling of the detection and electrophoresis systems. The close proximity of the working electrode to the exit of the separation channel results in symmetric peak shapes and efficient separations (50,000-100,000 plates/m). A direct comparison between the frequency-based electrochemical technique, sinusoidal voltammetry, and the more commonly used constant potential (DC) amperometry is made. Sinusoidal voltammetry is found to be roughly an order of magnitude more sensitive than DC amperometry, with calculated mass detection limits (S/N = 3) of 12 amol and 15 amol for dopamine and isoproterenol, respectively.     

Direct determination of carbohydrates, amino acids, and antibiotics by microchip electrophoresis with pulsed amperometric detection

The separation and detection of underivatized carbohydrates, amino acids, and sulfur-containing antibiotics in an electrophoretic microchip with pulsed amperometric detection (PAD) is described. This report also describes the development of a new chip configuration for microchip electrophoresis with PAD. The configuration consists of a layer of poly(dimethylsiloxane) that contains the microfluidic channels, reservoirs, and a gold microwire, sealed to a second layer of poly(dimethylsiloxane). Example separations of carbohydrates, amino acids, and sulfur-containing antibiotics are shown. The effect of the separation and injection potentials, buffer pH and composition, injection time, and PAD parameters were studied in an effort to optimize separations and detection. Detection limits ranging from 6 fmol (5 microM) for penicillin and ampicillin to 455 fmol (350 microM) for histidine were obtained.     

Improving the compatibility of contact conductivity detection with microchip electrophoresis using a bubble cell

A new approach for improving the compatibility between contact conductivity detection and microchip electrophoresis was developed. Contact conductivity has traditionally been limited by the interaction of the separation voltage with the detection electrodes because the applied field creates a voltage difference between the electrodes, leading to unwanted electrochemical reactions. To minimize the voltage drop between the conductivity electrodes and therefore improve compatibility, a novel bubble cell detection zone was designed. The bubble cell permitted higher separation field strengths (600 V/cm) and reduced background noise by minimizing unwanted electrochemical reactions. The impact of the bubble cell on separation efficiency was measured by imaging fluorescein during electrophoresis. A bubble cell four times as wide as the separation channel led to a decrease of only 3% in separation efficiency at the point of detection. Increasing the bubble cell width caused larger decreases in separation efficiency, and a 4-fold expansion provided the best compromise between loss of separation efficiency and maintaining higher field strengths. A commercial chromatography conductivity detector (Dionex CD20) was used to evaluate the performance of contact conductivity detection with the bubble cell. Mass detection limits (S/N = 3) were as low as 89 +/- 9 amol, providing concentration detection limits as low as 71 +/- 7 nM with gated injection. The linear range was measured to be greater than 2 orders of magnitude, from 1.3 to 600 microM for sulfamate. The bubble cell improves the compatibility and applicability of contact conductivity detection in microchip electrophoresis, and similar designs may have broader application in electrochemical detection as the expanded detection zone provides increased electrode surface area and reduced analyte velocity in addition to the reduction of separation field effects.     

Real-Time Imaging through Optical Fiber Array-Assisted Laser-Induced Fluorescence of Capillary Electrophoretic Enantiomer Separations

An advanced detection system based on laser-induced fluorescence imaging for capillary electrophoresis (CE) is presented. An optical fiber array was constructed for collection and transportation of the emitted fluorescent light to the charge-coupled device (CCD) camera. The fiber array makes the setup compact compared with a setup where the capillary is imaged through a camera objective. The imaging detector captures the sample zones in motion during the migration through the capillary. This allows unique studies on dynamic events otherwise unrevealed. During the study, unexplained nonlinear migration behavior was revealed. Enantiomer separations of dansylated amino acids using cyclodextrins, imaged between 1.5 and 12 cm of a 28-cm-long 50-μm i.d. capillary, were used for evaluation of the system. Comparing the optical fiber array with a camera lens system, the signal-to-noise-ratio (S/N) was 10 times higher. This is due to a combination of both higher signal and lower noise levels. To improve the S/N ratio further, a computer program for signal processing was designed. Using dichlorofluorescein, a concentration limit of detection (CLOD) of 350 pM was achieved and improved 10 times to 35 pM with computer postprocessing using 79 images. This is equal to 400 zeptomole for a 3-mm-long sample zone in a 50-μm i.d. capillary.     

Height measurement of microchip connecting pins by use of stereovision

A stereovision method for estimating the height of connecting pins on a microchip is described. The technique uses a centroidal method to simplify the calculation. A few seconds are required for a Pentium 586 PC to calculate the heights of 300 connecting pins on a 50 mm x 50 mm microchip. The method is described, and experimental results are presented. The optical system, which consists of two CCD cameras with long-focal-length lenses and a two-channel digital image grabber, is capable of in situ measurement.     

Interline Transfer CCD Camera for Gated Broadband Coherent Anti-Stokes Raman-Scattering Measurements

Use of an interline transfer CCD camera for the acquisition of broadband coherent anti-Stokes Raman-scattering (CARS) spectra is demonstrated. The interline transfer CCD has alternating columns of imaging and storage pixels that allow one to acquire two successive images by shifting the first image in the storage pixels and immediately acquiring the second image. We have used this dual-image mode for gated CARS measurements by acquiring a CARS spectral image and shifting it rapidly from the imaging pixel columns to the storage pixel columns. We have demonstrated the use of this dual-image mode for gated single-laser-shot measurement of hydrogen and nitrogen CARS spectra at room temperature and in atmospheric pressure flames. The performance of the interline transfer CCD for these CARS measurements is compared directly with the performance of a back-illuminated unintensified CCD camera.     

科学级CCD相机在星敏感器中的设计与应用

星敏感器通过探测天球上不同位置的恒星来确定航天器的姿态。星相机是星敏感器的成像系统。虽然近年来CMOS成像技术快速发展,但在科学级成像领域,特别是星敏感器应用中,CCD技术较成熟。本文的研究目的是研究一种探测能力强、数据更新快的用于星敏感器的成像系统。文中主要研究了基于TH7888A科学级CCD传感器的星相机的设计和应用,说明了CCD工作原理,详细分析了该种CCD传感器的星等探测灵敏度,论述了CCD星相机的设计方案。并用成像实验、动态范围测试、星等探测能力实验等实验验证了所设计的相机的性能。设计的星相机可以用给定的小型光学系统在60 ms以内的积分时间探测6等星,相机可达到10帧/秒的图像数据更新频率,满足短积分时间进行快速星光成像的要求。     

Chemiluminescence detection for hybridization assays on the flow-thru chip, a three-dimensional microchannel biochip.

Chemiluminescence (CL) detection is seldom used in two-dimensional solid support microarray platforms because adequate sensitivity and spatial resolution is difficult to achieve. The three-dimensional ordered microchannels of the Flow-thru Chip increase both the sensitivity and spatial resolution required for quantitative CL measurements on microarrays. Enzyme-catalyzed CL reactions for the detection of hybridizations on microchannel glass were imaged using a CCD camera. Signal uniformity, sensitivity, and dynamic range of the detection method were determined. The relative standard deviation of signal intensities across an array of 64 spots was 8.1%. A detection limit of 250 amol of target with a linear dynamic range of 3 orders of magnitude was obtained for a 3-h assay. Similar to two-color fluorescence measurements, multiple enzyme labels were employed to demonstrate two-channel chemiluminescence. A unique method for measuring the relaxation time of a chemiluminescent species is also described.     

数码相机CCD噪点的检测

CCD的噪点是影响数码相机成像质量的关键因素之一。本文讨论了CCD噪点形成的原因，提出了针对不同特性的噪点进行软件检测的两种方法，并对实验检测结果进行了分析。     

A two-dimensional X-ray diffraction pattern sensor using a solid state area sensitive detector

A novel system is described for the direct collection of two-dimensional X-ray diffraction patterns on a CCD video microchip and for the processing and storage of the collected data in a microcomputer. The system described is nearing completion; some of its characteristics and features are detailed. This system was designed for use with a limited area; it can, however, be easily extended to large area detection.