Method for detecting content of glutathione (GSH) in each cell by using microfluidic chip based laser induced fluorescence system

A technology of laser-induced fluorescence and glutathione content, applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of cumbersome detection operation, the sensitivity needs to be further improved, etc., to shorten the operation time, wide range of use, sensitivity high effect

Inactive Publication Date: 2015-06-24
SHANDONG UNIV
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Problems solved by technology

However, when this method detects substances in a single cell, it needs to undergo cell membrane breaking to detect intracellular substances, and the detection device uses an inverted microscope assembled with a photomultiplier tube. The detection operation is relatively cumbersome, and the detection sensitivity is still high. To be further improved

Method used

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  • Method for detecting content of glutathione (GSH) in each cell by using microfluidic chip based laser induced fluorescence system
  • Method for detecting content of glutathione (GSH) in each cell by using microfluidic chip based laser induced fluorescence system
  • Method for detecting content of glutathione (GSH) in each cell by using microfluidic chip based laser induced fluorescence system

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Embodiment 1

[0031] Example 1: A microfluidic-laser-induced fluorescence system for the detection of glutathione content in a single cell

[0032] Such as figure 1 with figure 2 As shown, the microfluidic-laser-induced fluorescence system for detecting glutathione content in a single cell includes a microfluidic chip 2, and the microfluidic chip 2 is a cross-shaped microfluidic chip, which is provided with a buffer Pool B, sample pool S, waste liquid pool SW and buffer waste liquid pool BW four liquid storage pools; the four liquid storage pools of the microfluidic chip 2 are respectively connected to the four power output terminals of the electric drive instrument 1; The microfluidic chip 2 is placed on the detection platform 3 of the laser-induced fluorescence system, and the detection platform 3 is connected to the data collector 4 for collecting the fluorescence emitted by the cells to be detected on the microfluidic chip after laser irradiation, The data collector 4 is connected wi...

Embodiment 2

[0034] Embodiment 2: Detection of GSH content in a single human red blood cell

[0035] Using the microfluidic-laser-induced fluorescence system of Example 1 to detect the GSH content in a single human red blood cell, the specific steps are as follows:

[0036] (1) Cell pretreatment: take 1.0ml of healthy adult blood and add K 2 EDTA anticoagulant in a 2ml plastic centrifuge tube, centrifuge at 1500rpm for 5min, and discard the upper layer solution. Add PBS to the remaining red blood cells to 1.0ml, mix gently and then centrifuge again, discard the supernatant. Repeat this 4-5 times until the supernatant is clear and transparent to prepare a red blood cell suspension, which is then counted with a hemocytometer. The low-concentration red blood cell suspension used was diluted from the high-concentration suspension.

[0037] (2) Drawing of glutathione standard curve: Before the experiment started, the microfluidic chip pipeline was washed with 0.1mol / L sodium hydroxide soluti...

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Abstract

The invention discloses a microfluidic chip based laser induced fluorescence system for detecting the content of glutathione (GSH) in each cell and a method for rapidly and continuously detecting the content of GSH in each cell by adopting the microfluidic chip based laser induced fluorescence system. After derivatization from 2,3-naphthalenedicarboxaldehyde (NDA) is carried out in a cell, the derivative generated from GSH and NDA in the cell can emit fluorescent light after being irradiated with laser. The sample injection, separation and detection are carried out in a microfluidic chip. Four power supply output ends of an electric driver are respectively connected a buffer tank B, a sample tank S, a solution waste tank SW and a buffer waste tank BW of the microfluidic chip. The microfluidic chip is arranged on the detection platform of the microfluidic chip based laser induced fluorescence system. The fluorescent light emitted by the cell is acquired by a data acquisition device, and then the fluorescent light is displayed by the software of a computer. The method disclosed by the invention can be used for detecting the content of GSH in each cell without the necessity of breaking the cell. The microfluidic chip based laser induced fluorescence system has high sensitivity and high resolution. By adopting the method and the microfluidic chip based laser induced fluorescence system, the operation difficulty can be decreased, and the operation time can be shortened.

Description

technical field [0001] The invention relates to a method for detecting glutathione content in a single cell by a microfluidic-laser-induced fluorescence system, and belongs to the technical field of biological detection. Background technique [0002] With the rapid development of technology and equipment in the field of life sciences, the research on cells has become more in-depth. The statistical average results obtained by cell population analysis conceal the differences between individual cells and cannot reflect the independent information of each cell. Cells are the basic unit of the structure and function of organisms. Rapid and sensitive detection of some important components in cells is of great significance for understanding many physiological and pathological processes and early diagnosis and treatment of diseases. [0003] Microfluidics is the integration of basic operating units such as sample preparation, reaction, separation, and detection in biological, chemi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 刘汝涛郝明路
Owner SHANDONG UNIV
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