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Online self-calibration laser induced fluorescence detection method based on electric charge coupling apparatus

A technology of charge-coupled device and laser-induced fluorescence, which is applied in the field of liquid chromatography and online self-calibration micro-analysis laser-induced fluorescence detection, which can solve the problems of difficult excitation light source and high price

Inactive Publication Date: 2005-03-16
SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the early application of PMT and the relatively mature technology, it has the widest application range. It is commonly used in literature reports and commercial detectors. The advantage is that the sensitivity is relatively high, but the price is high. The influence is very sensitive, so the detection system has to be completely protected from light or placed in a light-proof device, and the laser intensity is stabilized
Ordinary CCDs are mainly used to detect fluorescence imaging, or use the fluorescence intensity of the entire image as the detection object; the main advantage of ICCD is to improve the sensitivity to optical signals and the signal-to-noise ratio, but the application of these two optoelectronics in existing literature reports It is difficult to solve the problem of external stray light and unstable excitation light source when converting devices

Method used

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  • Online self-calibration laser induced fluorescence detection method based on electric charge coupling apparatus
  • Online self-calibration laser induced fluorescence detection method based on electric charge coupling apparatus
  • Online self-calibration laser induced fluorescence detection method based on electric charge coupling apparatus

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Effect test

Embodiment 1

[0036] Capillary Electrophoresis-Laser Induced Fluorescence Separation and Detection of Mixed Solutions of Methylene Blue and Nile Blue

[0037] Take a capillary of a certain length, burn a detection window of a suitable size, fix the capillary on the microscopic platform, adjust the microscopic platform, and place the detection window at an appropriate position in the optical path. Select the fluorescence detection area on the software interface, select the reference light area in the non-sample light area, and adjust other parameters of the program. The two ends of the capillary are immersed in the buffer solution, and connected to the two ends of the high-voltage power supply through platinum wire electrodes. Rinse the capillary with a buffer solution first, then replace the buffer solution at the injection end with the sample solution. After the sample is injected, replace the injection end with a buffer solution, apply a certain voltage for electrophoretic separatio...

Embodiment 2

[0046] Microchip Electrophoresis-Laser Induced Fluorescence Separation of Mixed Solutions of Methylene Blue and Nile Blue

[0047] Fix the microchip on the microscopic platform, adjust the microscopic platform, and set the detection window to the sample detection position in the light path. Select the fluorescence detection area on the software interface, and select the reference light area in the non-sample area, and adjust other parameters of the program. The buffer solution was manually dropped into the channel in advance, and connected to both ends of the high-voltage power supply through platinum wire electrodes. Rinse the separation channel with buffer solution first, and then drop the sample solution into the sampling tank. After electrosampling for a period of time, quickly rinse the sample solution in the sampling tank with buffer solution, and then drip the buffer solution into the sampling tank. Apply a certain voltage for electrophoretic separation, and a...

Embodiment 3

[0055] Microchip electrophoresis-semiconductor laser-induced fluorescence indirect detection of berberine

[0056] Operation is the same as embodiment two.

[0057] Laser: 630nm semiconductor laser; fluorescence collecting objective lens × 10 / 0.25, filter 720nm cut-off filter

[0058] Background buffer: 2.0*10 -5 M methylene blue (pH3.00, prepared in 5mM phosphate buffer)

[0059] Sample: 2.0*10 -5 M, prepared with pure distilled water

[0060] Injection: 0.50KV, 5.0s

[0061] Operation: 0.50KV, ~0.9μA

[0062] Total channel length: 50mm, effective separation length: 40mm

[0063] Under this condition, the minimum detection concentration of berberine is 2.0*10 -7 m

[0064] Microchip electrophoresis-laser-induced fluorescence diagram for the indirect detection of berberine with methylene blue Figure 5 shown.

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Abstract

It is an on-line self-correction laser leading fluorescence measurement method and its application in the analysis of chromatogram, capillary electrophoresis and core electrophoresis based on charge couple device. The method in this invention uses laser-leading sample to generate fluorescence characteristics and directly adopts CCD as measurement elements and translates the mock video signals collected by CCD into digital signals. It uses non-sample area light intensity in the video signals as reference signals when the self-correction video fluorescence measures the computer program and processes the video digital signals. It adopts reference correction method and eliminates the background interference signals in the sample light intensity and makes the recorded fluorescence signal data not sensitive to the around light interference and can reduce the background noisy interference signals without adding new hardware.

Description

technical field [0001] The invention relates to a laser-induced fluorescence confocal detection method, in particular to an online self-calibration micro-analysis laser-induced fluorescence detection method and system, and a liquid chromatograph, capillary electrophoresis instrument and chip used in conjunction with the detector Electrophoresis apparatus and detection method. Background technique [0002] Laser-induced fluorescence (LIF) detection is one of the most sensitive methods among all detection methods. The optical system of laser-induced fluorescence detection is divided into two types: non-confocal and confocal. Now, confocal laser-induced fluorescence detection is commonly used. The lasers used in commercial instruments are generally expensive, have a short service life, and are relatively complicated in structure and operation. Semiconductor lasers (or laser diodes) have quickly attracted people's attention for their advantages of low price, small size, long li...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N27/447G01N30/74G01N35/00
Inventor 许旭高红军乔善磊王前柴逸峰
Owner SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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