Chengke Xie

Development of an up-converting phosphor technology-based 10-channel lateral flow assay for profiling antibodies against Yersinia pestis

In this study, a 10-channel up-converting phosphor technology-based lateral flow (TC-UPT-LF) assay was developed to profile antibodies against Yersinia pestis. Ten expressed Y. pestis proteins were covalently conjugated with an up-converting phosphor particle to develop double-antigen sandwich immunochromatographic strips to detect corresponding antibodies. After optimization one by one, each strip was integrated into a TC-UPT-LF disc for simultaneously detection of different antibodies. A scanning biosensor was also developed to acquire the results. The performance of the TC-UPT-LF assay was evaluated by using standard samples and plague monkey serum samples. Fifty-one patient serum samples were detected by the TC-UPT-LF assay. The TC-UPT-LF disc could be stable for 10 days at 37°C with an average CV of 10.3%. Its sensitivity and qualitative results are comparable to those of ELISA. Its linearity fitting coefficient of determination (R2) for different antibody detection is between 0.93 and 0.99. Besides F1 antibody, the LcrV and YopD antibodies also showed higher positive ratio than the other seven antibodies, as 100% (13/13) and 92% (12/13) in monkey sera and 86.3% (44/51) and 66.7% (34/51) in patient sera, respectively. It is suggested that the TC-UPT-LF assay has been successfully developed for multi-detection and LcrV and YopD can be the potential diagnostic markers of the plague.

A Simple Optical Reader for Upconverting Phosphor Particles Captured on Lateral Flow Strip

A simple optical reader was developed for rapid and sensitive quantification of lateral flow (LF) strip with upconverting phosphor (UCP) particles as reporters. The excitation beam from a 980 nm laser diode (LD) was focused to a 4 mm (length) times 0.02 mm (width) rectangular spot on LF strip. A rectangular slit with 4 mm times 0.02 mm before the photomultiplier tube (PMT) which was selected as receiver was introduced as field diaphragm to prevent stray light from entering the PMT. The scanning resolution was 20 mum, and the scanning speed was 0.75 mm/s, and the scanning range was 10 mm. With the above optical reader, serial dilutions of Yersinia pestis F1 antigen were detected to achieve the test sensitivity was 5 ng/ml, and the dynamic range reached 150 ng/ml. The algorithms of the low-pass filter and the first derivative were used to search the boundaries of T line and C line from the original data acquired by the reader. A four-parameter logistic mathematical model was used to deduce the quantitative equation for determination of unknown F1 antigen concentration. Our 1-D optical reader possessed the following characteristics: high 1-D testing resolution, high sensitivity, simple structure, simple data processing method, high testing efficiency, and small total volume.

Theoretical model and quantif ication of reflectance photometer

The surface morphology of lateral flow (LF) strip is examined by scanning electron microscope (SEM) and the diffuse reflection of porous strip with or without nanogold particles is investigated. Based on the scattering and absorption of nanogold particles, a reflectance photometer is developed for quantification of LF strip with nanogold particles as reporter. The integration of reflection optical density is to indicate the signals of test line and control line. As an example, serial dilutions of microalbunminuria (MAU) solution are used to calibrate the performance of the reflectance photometer. The dose response curve is fitted with a four-parameter logistic mathematical model for the determination of an unknown MAU concentration. The response curve spans a dynamic range of 5 to 200 µg/ml. The developed reflectance photometer can realize simple and quantitative detection of analyte on nanogold-labeled LF strip.

Measurement precision improvement for excimer laser pulse energy sensor based on a special finite impulse response filter

In deep ultraviolet lithographic tools, precise measurement of excimer laser pulse energy is essential. However, system noise of the energy sensor, which is used to monitor the laser pulse during exposure dose control, complicate pulse energy measurement. To improve measurement precision, a specific finite impulse response filter is proposed for signal denoising. Particularly, the convolution kernel of the specific finite impulse response filter is derived from the nonlinear fitting of the energy sensor output. Experimental results show that the measurement precision of the energy sensor improves 2-3 times with the specific finite impulse response filter. To verify the effectiveness of the energy sensor, it is used for transmittance measurement; the measurement result of a Corning 7980 witness sample is consistent with that measured by a commercial ultraviolet spectrophotometer and the measurement accuracy is 0.11% (k = 2).

Accurate measurement of illumination uniformity using spot sensor based 2-dimension scanning method

Illumination uniformity of the illuminator is significant for achieving stringent critical dimension (CD) control for lithography machine. In order to achieve high uniform illuminating, there is an urgent demand of accurate measurement for illumination uniformity. The difficulty for accurate measurement of illumination uniformity for the illuminator mainly lies in two aspects: the illumination plane is large; excimer laser pulse energy is variable from pulse to pulse. In this work, a spot sensor based 2-dimension scanning method for illumination uniformity measurement is proposed, where the spot sensor in combination with a 2-dimension moveable stage is located in the illumination plane of the illuminator is used to scan the illumination plane point-by-point so as to obtain the whole irradiation distribution. To improve measurement accuracy, the energy sensor of the illuminator serves as the reference and monitors each pulse in real-time showing benefit of excimer laser pulse energy fluctuation eliminating. Secondly, the used spot sensor is modified for strict synchronization control of the spot sensor and the energy sensor so that the measurement precision can be improved. Measurement results show that X direction transient non-uniformity, Y direction transient non-uniformity and X direction integral non-uniformity of the illuminator are 5.14%, 5.55% and 2.16% respectively with a measurement uncertainty of 0.31% (k=2). It is proved that the proposed method is effective and helpful for further system optimizing and alignment.

Scattering performance analysis of reflectance photometer for quantifying gold labeled test strips

Gold labeled immunochromatography assay is widely used in many fields. Quantitative test can be realized by using a reflectance photometer. However, theoretical analysis of the strip and the photometer has seldom been reported. In this paper, the microstructure of immunochromatographic strip labeled by nanogold particles is analyzed with scanning electron microscope (SEM). Based on the SEM images of the strip, Mies scattering theory is used to investigate the scattering behaviors of particles in the strip, and Lambert cosine law is applied to analyze the diffuse reflection of porous strip. Besides, a reflectance photometer for gold labeled test strip has been developed for fast quantification of immunochromatography assay in our group. Theoretical model is achieved by introducing the parameter of the developed reflectance photometer. The calculated scattering signal distribution is well consistent with that measured by the reflectance photometer.

Improvement of up-converting phosphor technology-based biosensor

A novel biosensor based on up-converting phosphor technology (UPT) was developed several years ago. It is a kind of optical biosensor using up-converting phosphor (UCP) particles as the biological marker. From then on, some improvements have been made for this UPT-based biosensor. The primary aspects of the improvement lie in the control system. On one hand, the hardware of the control system has been optimized, including replacing two single chip microcomputers (SCM) with only one, the optimal design of the keyboard interface circuit and the liquid crystal module (LCM) control circuit et al.. These result in lower power consumption and higher reliability. On the other hand, a novel signal processing algorithm is proposed in this paper, which can improve the automation and operating simplicity of the UPT-based biosensor. It has proved to have high sensitivity (~ng/ml), high stability and good repeatability (CV<5%), which is better than the former system. It can meet the need of some various applications such as rapid immunoassay, chemical and biological detection and so on.