Photothermal Detection of Individual Gold Nanoparticles: Perspectives for High‐Throughput Screening

We use photothermal microscopy to detect and image individual gold nanoparticles that are either embedded in a polymer film or immobilized in an aqueous environment. Reducing the numerical aperture of the detection optics allows us to achieve a 200‐fold‐enlarged detection volume while still retaining sufficient detectivity. We characterize the capabilities of this approach for the detection of gold colloids with a diameter of 20 nm, with emphasis on practical aspects that are important for high‐throughput‐screening applications. The extended detection volume in combination with the stability of the photothermal signal are major advantages compared to fluorescence‐based approaches, which are limited by photoblinking and photobleaching. Careful consideration is given to the trade‐off between the maximum increase in local temperature that can be tolerated by a biological specimen and the minimum integration time needed to reliably determine whether a given volume contains a target species. We find that our approach has the potential to increase the detection‐limited flow rate (i.e. the limit given by the detection volume divided by the minimum detection time) by two to three orders of magnitude.     

Attogram detection limits using laser induced fluorescence

Laser induced fluorescence has produced outstanding detection limits in liquid phase analysis. This paper presents a general method for optimizing detection limits as a function of sample volume.     

Rapid microRNA detection using power-free microfluidic chip: coaxial stacking effect enhances the sandwich hybridization

We present a new method for rapid microRNA detection with a small volume of sample using the power-free microfluidic device driven by degassed PDMS. Target microRNA was detected by sandwich hybridization taking advantage of the coaxial stacking effect. This method allows us to detect miR-21 in 20 min with a 0.5 μL sample volume at a limit of detection of 0.62 nM. Since microRNAs can act as cancer markers, this method might substantially contribute to future point-of-care cancer diagnosis.     

基于气液相化学发光技术的臭氧在线检测方法

基于鲁米诺(luminol) 化学发光体系,采用自主研发的在线臭氧浓度检测仪,建立了一种实时在线检测臭氧浓度的方法,用于分析测定痕量浓度水平的臭氧气体。 考察了鲁米诺、氢氧化钾、部分醇类化合物和表面活性剂等因素对化学发光强度的影响。 结果表明,在鲁米诺(0.005 mol/L)、氢氧化钾(0.05 mol/L)体系中加入乙二醇(体积分数1.5%)、甲醇(体积分数1.5%)、乙醇(体积分数1.0%)、丙三醇(体积分数3.0%)能显著增强鲁米诺体系检测O₃的化学发光信号,而甲醛溶液 (体积分数3.0%)能有效抑制NO₂信号的干扰。 同时,测得检测臭氧的检出限为1.26 μg/m³、相对标准偏差为0.32%,相对误差为0.75%。 利用该体系测定臭氧,具有信号稳定、精密度好、准确度高、检出限低等优点,适用于大气中微量O₃的在线连续检测。     

Sensitive and Accurate Determination of Cobalt at Trace Levels by Slotted Quartz Tube-Flame Atomic Absorption Spectrometry Following Preconcentration with Dispersive Liquid–Liquid Microextraction

Slotted quartz tube-flame atomic absorption spectrometry (SQT-FAAS) was used as a sensitive technique for the determination of cobalt, an element has toxic effects on living organisms at high doses. For the preliminary preconcentration of cobalt prior to the analysis, dispersive liquid–liquid microextraction (DLLME) was used. The instrument is equipped with a SQT to further increase the sensitivity by increasing the residence time of cobalt atoms in the light path emitted by a hollow cathode lamp. In the complex formation step, pH, the volume of buffer solution, the concentration of 1,5-diphenylcarbazone, and the volume of ligand were optimized. In addition, all of the system parameters, including the type and volume of the extraction solvent, dispersant type and volume of solvent, type of salt and the volume for the dispersion liquid–liquid microextraction, were optimized to obtain the lowest detection limit. Under the optimum conditions, the detection power of FAAS was improved by a factor of 86.56 fold using DLLME-SQT-FAAS. The limit of detection for the DLLME-SQT-FAAS system was 0.97?µg L^?1. The applicability of the developed method was verified in tap and waste water samples by spiking measurements. The percentage recovery values for these were determined to be 91.7% and 111.0%, respectively.     

Optimisation design of cylindrical containers for improving the detection efficiency of a high-purity germanium detector using the LabSOCS

The LabSOCS software was used to optimise the geometric dimensions of cylindrical containers and improve the detection efficiency of a high-purity germanium detector for the measurement of environmental radioactivity samples. The optimum ratio of diameter and height for different sample volumes were determined. In addition, the degrees of influence of the sample volume and density when the amount of sample was fixed were compared. Results show that the effect of sample volume on detection efficiency was significantly greater than that of sample density for a given sample amount.     

Electrochemical detection of amino acids based on cucurbit[7]uril-mediated three-dimensional gold nanoassemblies

In this paper, cucurbit[7]uril (CB[7])-mediated three-dimensional gold nanoassemblies were successfully prepared to increase the loaded amount of CB[7] and enhance the electrochemical detection of amino acids. Particle sizes of gold nanoparticles (AuNPs) significantly affect stability and detection sensitivity of nanoassemblies. The volume of gold nanoassemblies first increased and then decreased with the increase of CB[7] concentration. The 3D gold nanoassemblies composed of 16 nm AuNPs and 100 µmol/L CB[7] had excellent stability and maximum volume, exhibiting more sensitive detection for a variety of amino acids. And the detection limits of aromatic amino acids are lower in virtue of the higher binding constant between aromatic amino acids and CB[7]. This study will develop and deepen our understanding of molecular recognition in amino acids detection.     

Amperometric detection for capillary flow injection analysis

Ultrasmall-volume measurements of oxidizable compounds have been accomplished by coupling a capillary flow injection system with amperometric detection. Remarkably low (femtomole) mass detection limits result from the combination of nanoliter sample volume and the inherent sensitivity of the wall-jet detector. A substantial economy of reagent consumption and disposal accrues from the operation of the nl/min flow regime. Variables influencing the physical dispersion in the capillary flow injection system, including capillary length, sample volume or flow rate, are explored and optimized.     

单分子流式检测仪的研制

采用激光诱导荧光和流体动力学聚焦技术成功地研制出单分子流式检测仪, 实现了对水溶液中单个藻红蛋白及单个DNA分子片段的检测, 检测速率可达到每秒几十次. 与单分子荧光显微术相比, 流式分析将固定的标本台改为流动的单分子悬液, 大大提高了检测速率和统计精确性, 更加适合生物样品的快速、超高灵敏分析.     

Detection limits of thermal conductivity and photoionization detectors in high speed narrow bore CGC

Not only the required input band width and the availability of compatible instrumentation limit the reduction of column diameter but so do the column sample capacity and detector characteristics, such as volume and sensitivity. In this paper the scope and limitations of thermal conductivity and photoionization detection at atmospheric and reduced pressures for capillary gas chromatography are discussed at length. It is shown that the sensitivity of a thermal conductivity detector is inversely proportional to the pressure. Reduction of cell volume and decrease of pressure appear equivalent with respect to minimum detectable amount and peak broadening. This results in femtogram detection limits for column diameters as low as 10 μm. For photoionization detectors the sensitivity did not improve at reduced pressures, so that the effect of reduced pressure and addition of make-up gas on the detection limits is the same. Nevertheless, the applicability of a low volume photoionization detector (40 μl) for capillary columns with a diameter as low as 50 μm will be demonstrated by series coupled PID and FID detectors and illustrated with various applications.     

Pore size distribution measurements in small samples and with nanoliter volume resolution by NMR cryoporometry

NMR cryoporometry is demonstrated to provide access to pore size and pore size distribution in small amounts of porous materials. With a conventional NMR spectrometer and probe, the detection limit for total pore volume is shown to be on the order of 100 nL with a volume resolution on the order of a few nanoliters.     

On-line preconcentration methods for capillary electrophoresis

The limits of detection (LOD) for capillary electrophoresis (CE) are constrained by the dimensions of the capillary. For example, the small volume of the capillary limits the total volume of sample that can be injected into the capillary. In addition, the reduced pathlength hinders common optical detection methods such as UV detection. Many different techniques have been developed to improve the LOD for CE. In general these techniques are designed to compress analyte bands within the capillary, thereby increasing the volume of sample that can be injected without loss of CE efficiency. This on-line sample preconcentration, generally referred to as stacking, is based on either the manipulation of differences in the electrophoretic mobility of analytes at the boundary of two buffers with differing resistivities or the partitioning of analytes into a stationary or pseudostationary phase. This article will discuss a number of different techniques, including field-amplified sample stacking, large-volume sample stacking, pH-mediated sample stacking, on-column isotachophoresis, chromatographic preconcentration, sample stacking for micellar electrokinetic chromatography, and sweeping.     

Improving process methodology for measuring plutonium burden in human urine using fission track analysis

The aim of this paper is to clearly define the chemical and nuclear principles governing Fission Track Analysis (FTA) to determine environmental levels of²³⁹Pu in urine. The paper also addresses deficiencies in FTA methodology and introduces improvements to make FTA a more reliable research tool. Our refined methodology, described herein, includes a chemically-induced precipitation phase, followed by anion exchange chromatography and employs a chemical tracer,²³⁶Pu. We have been able to establish an inverse correlation between Pu recovery and sample volume and our data confirms that increases in sample volume do not result in higher accuracy or lower detection limits. We conclude that in subsequent studies, samples should be limited to approximately two liters. The Pu detection limit for a sample of this volume is 2.8 μBq/l.     

Calculation Methods of Solution Chemical Potential and Application in Emulsion Microencapsulation

Several new biased sampling methods were summarized for solution chemical potential calculation methods in the field of emulsion microencapsulation. The principles, features, and calculation efficiencies of various biased Widom insertion sampling methods were introduced, including volume detection bias, simulation ensemble bias, and particle insertion bias. The proper matches between various types of solution in emulsion and biased Widom methods were suggested, following detailed analyses on the biased insertion techniques. The volume detection bias methods effectively improved the accuracy of the data and the calculation efficiency by inserting detection particles and were suggested to be used for the calculation of solvent chemical potential for the homogeneous aqueous phase of the emulsion. The chemical potential of water, argon, and fluorobenzene (a typical solvent of the oil phase in double emulsion) was calculated by a new, optimized volume detection bias proposed by this work. The recently developed Well-Tempered(WT)-Metadynamics method skillfully constructed low-density regions for particle insertion and dynamically adjusted the system configuration according to the potential energy around the detection point, and hence, could be used for the oil-polymer mixtures of microencapsulation emulsion. For the macromolecule solutes in the oil or aqueous phase of the emulsion, the particle insertion bias could be applied to greatly increase the success rate of Widom insertions. Readers were expected to choose appropriate biased Widom methods to carry out their calculations on chemical potential, fugacity, and solubility of solutions based on the system molecular properties, inspired by this paper.     

Analysis of the influence of interdetector volume on local calibration in size exclusion chromatography with dual multiangle-light-scattering/concentration detection

A comparison of the s- and w-detections of molecular weight, a necessary condition for precise determination of interdetector volume in size exclusion chromatography of polymers with the dual light scattering--concentration detection, is examined. From the theoretical analysis for a polymer with the log-normal molecular weight distribution it follows that the error in determination of the local calibration, i.e., of a logarithmic dependence of molecular weight on elution volume obtained by the s-detection, from the known dependence of radius of gyration on elution volume and of molecular weight on the radius of gyration, remains linear and is slightly shifted with respect to that obtained by the w-detection (dual detection giving the weight-average molecular weight) towards higher molecular weights, the difference being below the experimental error. It also follows that the error in the slope of local calibration found by the w-detection is given by the ratio of the error in interdetector volume and the variance of the elution curve. This is demonstrated on several polymer samples with symmetric elution curves of polymer samples differing in polydispersity indices. The range of this rule depends on the broadness and symmetry of elution curves. The precision of the interdetector volume determination depends therefore strongly on the Mw/Mn ratio of the sample.     

Application of time-resolved fluorescence for imaging-based multisample and multianalyte detection in single microtiter wells

Enzyme-linked immunosorbent assays are routinely used in laboratories around the world and ensure highly specific protein detection. Often, more than one analyte needs to be determined in a single sample and numerous protein arrays for multianalyte detection of a single sample have been developed to address this problem. They have the potential to analyze several dozen or even more analytes in an assay volume of usually around 100 μL. However, due to the presence of numerous different antibodies, these multianalyte sandwich immunoassays suffer from undesired cross-reactivities between the antibodies which lead to a loss of assay specificities. Here, we present an assay principle which allows, e.g., a detection of an analyte in a sample volume of only 1 μL in a normal 96-microtiter well plate, so that up to 100 analytes can be determined from a 100 μL sample volume, but in separate wells. This eliminates antibody cross-reactivities. The assay is based on the biotinylated time-resolved fluorophore EuLH used as a PEG₁₁-dye conjugate in combination with ExtrAvidin® to ensure high signal-to-background ratios. The model protein epidermal growth factor (EGF) was detected with the established sandwich immunoassay and showed assay parameters comparable to commercially available ones. Furthermore, the assay principle enables a spatial resolution of the assay signal. Here, we demonstrated the application of the new detection system for universal imaging-based analysis of individual spots in one single 96-microtiter well by applying it to multisample and also multianalyte detections. In the case of the multisample analysis approach, a considerable reduction of the required sample volume to only 1 μL in a single 96 microtiter well could be achieved.     

Integrated microfluidic UV absorbance detector with attomol-level sensitivity for BSA

An integrated UV absorbance detection system employing a novel silicon-in-plastic technology to seamlessly integrate bare UV photodiode chips into polymer microfluidic systems has been developed. Detection platforms fabricated using this approach exhibit exceptionally low concentration and mass detection limits down to 15 nM and 9.8 amol, respectively, for bovine serum albumin (BSA) as a model protein. In addition to providing high sensitivity, sub-nanoliter detection volumes are enabled by the use of direct photodiode integration. The fabrication methodology is detailed, and system performance metrics including detection limits, detection volume, dynamic range, and linearity are reported.     

Micro-impedance cytometry for detection and analysis of micron-sized particles and bacteria

The sensitivity of a microfluidic impedance flow cytometer is governed by the dimensions of the sample analysis volume. A small volume gives a high sensitivity, but this can lead to practical problems including fabrication and clogging of the device. We describe a microfluidic impedance cytometer which uses an insulating fluid to hydrodynamically focus a sample stream of particles suspended in electrolyte, through a large sensing volume. The detection region consists of two pairs of electrodes fabricated within a channel 200 μm wide and 30 μm high. The focussing technique increases the sensitivity of the system without reducing the dimensions of the microfluidic channel. We demonstrate detection and discrimination of 1 μm and 2 μm diameter polystyrene beads and also Escherichia coli. Impedance data from single particles are correlated with fluorescence emission measured simultaneously. Data are also compared with conventional flow cytometry and dynamic light scattering: the coefficient of variation (CV) of size is found to be comparable between the systems.     

Integrated microsystem for dielectrophoretic cell concentration and genetic detection

We have directly integrated a continuous-flow, electrokinetic method of bacterial cell concentration with room temperature, sequence-specific genetic detection. The system we have developed traps cells from a continuous-flow sample stream via dielectrophoresis, providing a 160-fold increase in cell concentration. PDMS microvalves then define a 100 nL volume around the trapped cells to which cell lysis buffer and genetic detection components are introduced. Direct, optical detection of Escherichia coli MC1061 cells is then accomplished via the sequence-specific hybridization of an rRNA-directed optical molecular beacon. This integrated microsystem is capable of sequence-specific genetic detection of 25 cells within 30 min.     

Toward sensitive, quantitative point-of-care testing (POCT) of protein markers: miniaturization of a homogeneous time-resolved fluoroimmunoassay for prostate-specific antigen detection

Point-of-care testing (POCT) systems which allow for a sensitive, quantitative detection of protein markers are extremely useful for the early detection and therapy progress monitoring of cancer. However, currently commercially available POCT devices are mainly limited to the qualitative detection of protein markers. In this study we demonstrate the successive miniaturization of a sensitive and fast assay for the quantitative detection of prostate-specific antigen (PSA) using a well established and clinically approved homogeneous time-resolved fluoroimmunoassay technology (TRACE?) on a commercial plate-reader system (KRYPTOR?). Regarding the initial requirements for the development of POCT devices we applied a 30-fold assay volume reduction (150 μL to 5 μL) to achieve a reasonable lab-on-a-chip volume and a 24-fold and 120-fold excitation pulse energy reduction to achieve reasonable pulse energies for low-cost miniature excitation sources. Due to highly efficient optimization of key POCT parameters our miniaturized PSA assay achieved a 30% increased sensitivity and a 2-fold improved limit of detection compared to the standard plate-reader method. Our results demonstrate the successful implementation of key parameters for a significant miniaturization and for cost reduction in the clinically approved KRYPTOR? platform for protein detection. The technological alterations required are easy-to-implement and can be immediately adapted for more than 30 diagnostic protein markers already available for the KRYPTOR? platform. These features strongly recommend our assay format to be utilized in innovative, sensitive, quantitative POCT of protein markers.