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Microfluidic chip detection of single cells O_2 ~ ( - ) study

Author: NieSuJuan
Tutor: XuKeHua；TangBo；LiQingLing
School: Shandong Normal University
Course: Analytical Chemistry
Keywords: Microfluidic Chips Gantry injection pinched injection combination Laser induced fluorescence detection Superoxide
CLC: Q813
Type: Master's thesis
Year: 2011
Downloads: 66
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Abstract

The cell is the basic unit of biological structures and activities of life. Explore the law and the essence of the life activity of organisms, it is necessary to study the cells based. The analysis of groups of cells is easy to overlook the differences between the cells, single-cell analysis is to analyze the chemistry, biology and medicine penetration between the development of the interdisciplinary field, can be a good solution to this problem. In particular, the quantitative detection of chemical components within a single cell, for the understanding of basic cellular functions, both inside and outside the cell contact, and a small amount of abnormal cells detect and identify a large number of cell populations are of great benefit. Superoxide is a precursor of other reactive oxygen once the excess, not only the large number of biological molecules have toxic effects, but also transforming into a more toxic radicals such as hydroxyl radical, hydrogen peroxide, etc.. Therefore, the detection of superoxide is of great significance for the study of cell metabolism and cell toxin pathogenesis. Microfluidic chip, also known as micro total analysis systems or lab-on-a-chip, 1990 by Manz and Widmer. In recent years, the microfluidic chip single-cell research concern, because it has its channel size and cell diameter, and less reagent consumption, rapid analysis, etc.. The goal of the development of microfluidic chip miniaturization, integration, automation and simplification. Although the integrated micro-pumps and micro-valves microfluidic system, used in single-cell component analysis, However, due to the micron chip channel, its establishment is still quite difficult. Simple Circuit component analysis of single cells is usually achieved by the combination of static pressure and the electric force or the electrostatic force generated by the type of liquid level Poor accompanied injector typically by simple injection, pinched injection and Doors type injection. The three injector injection the cells, cell loading, the cells captured by the two-step operation is completed Further, since the size of the micron-level passage, the cells exactly is controlled to have a certain degree of difficulty in the intersecting position of the chip channel. In short, the above-mentioned method for single cell analysis step tedious, time-consuming. Microchip detection of superoxide is just a story, its manipulation means of a combination of static pressure and electric force, injection mode pinched injection the probe selectivity dihydro ethidium (DHE). Detection of superoxide in a single cell-based the microfluidic chip gantry injection with pinched injection method of combining the high selectivity of 2 - chloro -1,3 - diphenyl thiazoline cyclohexane as a probe, (DBZTC) to solve the lack of the above methods of operation, mainly to carry out the research work of the following two parts: (a) based on the the microfluidic chip electric gantry injection (loading phase) with pinched injection (separation stage ) a combination of methods to detect the single HCC intracellular superoxide anion. Room synthetic probe 2 - chloro-1 ,3 - dibenzothiophene the oxazoline cyclohexane (DBZTC) on specific reaction with superoxide. Manifold diagram of the the superoxide oxidation products (DBO) to prove the injection phase of the gantry injection mode (loading phase). Cell injection, cell loading, cell capture step automated. Cells in the 15s was quickly captured. Cell activity experiments prove that the electric field intensity is appropriate injector. The DBO linear range of 2.1-102.8 amol, limit of detection (S / N = 3) 0.82 amol (8 nM). The average single cell content of superoxide and separation efficiency were 6.22 ± 3.49 amol and 4.73 × 104 number of theoretical plates. This method to speed up the progress of the single-cell analysis. (B) combined with electric gantry-based microfluidic chip static pressure injection (loading phase) (separation stage) pinched injection method for the detection of superoxide anion in a single liver cancer cells. Our laboratory synthesis of 2 - chloro-1 ,3 - dibenzothiophene the oxazoline cyclohexane (DBZTC) as a fluorescent probe. The fluorescent dye rhodamine 6G manifold that the injection phase (loading phase) for the gantry injection mode. A wide range of cell densities (1 × 10 ~ 5-1.0 × 10 ~ 6 cells? ML), the injection time is short (4s). Cell injection, cell loading, cell capture step is completed. DBO linear range of detection limit (S / N = 3) were 2.1-103 amol and 1.0 amol. The superoxide content within the average individual cells was 7.82 ± 3.87 amol. This method is simple, fast and accurate.

Microfluidic chip detection of single cells O_2 ~ ( - ) study

Abstract

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