In order to solve the problems of many defects in common digital PCR chips, an integrated droplet digital PCR chip was designed based on microfluidic, biology, optoelectronics, machinery and computer technology, which integrates droplet generation part, PCR amplification part and droplet fluorescence detection part. The droplet generation part used the flow focus micro structure to achieve droplet generation with two phase liquid, where the oil was used as continuous phase, the sample liquid as discrete phase. After the droplet generation process, the droplets were injected into collection chamber by the pressure of the pump. The amplification part uses the flat-plate PCR to achieve nucleic acid amplification in the integrated droplet digital PCR chip. After amplification, the integrated droplet digital PCR chip moves to detection location, under the irradiation of the excitation light the specificity target in the droplets will emit fluorescence, thus the fluorescence image is scanned by COMS camera and the result was analyzed by the software. The chips were made through injection molding process and the autofluorescence intensity of chip material was weak. Experimental results show that the coefficient of variation (CV) value of the droplet diameter was less than 2% and the temperature uniformity of the chip was less 0.5°C. That proves the chip designed has good droplet formation and thermal conductivity for nucleic acid amplification. The chip designed can reduce the manual operation error, operation difficulty and avoid the problems of droplet fusion, breakage and sample pollution in the process of pipetting.
In order to ensure the detection accuracy of laser-induced fluorescence detection, the micro droplet digital PCR material was required to have weak autofluorescence. The autofluorescence characteristic of micro droplet digital PCR chip was studied in this paper. Firstly, the chip was designed according to the principle of micro droplet digital PCR. Cycloolefin copolymers (COP) and cycloolefin polymers (COP) in organic polymers were selected as the materials of the chip because of their excellent biological characteristics. The laser induced fluorescence device was build. The excitation laser wavelength were FAM (Emission band:522nm, Excitation band:470nm) and HEX (Emission band:553nm, Excitation band:530nm) respectively. The fluorescence image of chip under different exposure time and different band was extracted by COMS camera. And read the fluorescence value through the computer software ImageJ. The glass plate was used as the reference for autofluorescence measurement of chip. From the test results, COC chip showed a good autofluorescence property compared with COP chip, which has lower autofluorescence value under laser excitation. Therefore, for the quantitative detection of trace nucleic acid, the high fluorescent background noise of COP chip will increase the threshold of negative positive discrimination which cloud lead to discrimination error and influence the requirements of detection accuracy. The research results provide a basis for the selection of high-precision micro droplet digital PCR chip materials.
With the continuous impact of COVID-19, the demand for rapid genetic diagnosing at the inspection and quarantine site, emergency treatment of sudden infectious diseases and clinical in vitro diagnosis was increasing rapidly. In order to achieve rapid, automatic nucleic acid extraction and detection, an automatic diagnostic system which integrates nucleic acid extraction, amplification and biochip fluorescence detection is designed. The diagnostic system designed based on multidisciplinary intersection of biology, optoelectronics, machinery and computer technology. At first, the nucleic acid extraction part has 1-8 sample flux and uses the principle of the silicon filter can adsorb nucleic acid at different pH to achieve nucleic acid extraction. After the extraction process, the nucleic acid is injected into biochip through robotic arm. There is a specific microarray reaction chamber on the chip, which can be combined with a specific biological substance; Secondly the amplification part used the principle of flat-plate PCR to achieve nucleic acid amplification in the biochip; After amplification, the nucleic acid detection realized under the irradiation of the excitation light at 530nm, the specificity target on the biochip will emit light, thus the fluorescence image scanned by COMS camera and the result analyzed by the software on PC; At last device control, automation and detection of the integrated device realized by the STM32 single-chip microcomputer and CAN communication method to establish a device communication network. The diagnostic system will automate workflow, and provides a result that reports on the detection and interpretation of targets in the samples. The imaging module resolution was less than 10 microns/pixel. The CV value of the sample was less than 10%. The results provide that the diagnostic system can provide more accurate and more automated equipment for nucleic acid extraction, amplification and detection.
Gene biochip imaging and scanning technology is a more advanced gene diagnosis technology after fluorescence quantitative PCR technology, second-generation sequencing technology and digital PCR technology. Due to the great influence of fluorescence microscopy system, the imaging and scanning technology of gene biochip with high imaging efficiency and accuracy has become a hot topic widely concerned by scholars. In this paper, two-channel laser was used as the excitation light source of the biochip, and the fluorescence imaging system of the switchable fluorescence filter was designed to realize the detection of FAM and HEX biological fluorescence signals; According to the working principle of the gene biochip imager, the finite element analysis method is used to analyze the thermal structure of the optical system, and the internal structural parameters of the optical system are optimized according to the simulation results. According to the optimization result, the illumination time of each object pixel is controlled to utilize the fluorescence information more efficiently, and the fluorescent bleaching effect is reduced without sacrificing the image quality, and the PCR amplification oil droplet quantitative reading and display is performed in real time. The gene biochip imaging system proposed in this paper can be used to image, obtain, read, process and display the detected chip with a resolution of 10 m/ pixel, a minimum detection limit of < 10 fluorescent molecules/square micron, and a sample detection repeatability of < 10%.The results show that the gene biological chip imaging system designed by this research has the advantages of low noise, low cost and fast scanning speed, and provides a theoretical basis for precision medical fields such as early diagnosis and cure evaluation of clinical tumors. The results show that the gene biochip imaging system designed by this research has the advantages of low noise, low cost and fast scanning speed, and provides a theoretical basis for precision medical fields such as early diagnosis and cure evaluation of clinical tumors.
The intelligent control of simulation target with infrared imaging target in the indoor and outdoor
environment can effectively and quantitatively evaluated the parameters such as the minimum resolution
temperature difference(MRTD)and spatial resolution of airborne forward looking infrared, infrared detection
and tracking, infrared alarm, and etc.
This paper focused on introducing the working principles of the intelligent control simulation target of
Infrared imaging target, studying the thermal radiation characteristics of the infrared target surface material,
analyzing the influences of the infrared radiation energy distribution, and developing the intelligent control
simulation target with IR imaging target for hardware-in-the-loop simulation test. The intelligent control
simulation target which area was 5 ㎡ and concluded 44 infrared targets including two kinds of infrared
targets ,0.25m×0.25m, and 0.25m×0.5m, achieved 1℃~10℃ temperature simulation of target and the
background, and temperature control precision better than 0.5℃. Field test requirements were achieved by
actual test.
In order to provide a set of field test equipment for the infrared system of modern weapon equipments and other optics instruments, a set of large-scale resistance-type infrared target system was designed. First, the large-scale infrared target was designed in modular construction. It was decomposed into several independent and controllable units. Then the working principle of the system was introduced. Three modes of thermal exchange (conduction, convection and radiation) and the computing methods for each mode were given under thermal equilibrium conditions through modeling and simulating. Periphery electro-circuit and control software were carried out as well. Finally, the performance of the system was tested. Meanwhile novel ways of temperature compensation to improve the uniformity of the surface temperature of the target was introduced. The experimental results show that the infrared target could meet test requirements for infrared imaging weaponry which wavelengths from 8 μm to 14 μm. The temperature control precision can reach 0.5ms. In conclusion, the infrared target system can satisfy requirements of reliability, high precision, as well as strong anti-jamming and stabilization.
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