| Na+, K+, Ca2+, Mg2+are four intracellular important metal ions, and their levels areclosely related of cellular physical and pathological processes. Wherein, Na+and K+areinvolved in regulation of important electrical physiological processes, such as cell mem-brane potential, and osmotic pressure, and their abnormal content changes will induce arr-hythmias and cardiovascular dysfunction and other diseases. Ca2+is the basis for the nor-mal physiological function of cell proliferation, division, movement, energy metabolism,oxygen metabolism, cell membrane permeability and stability, messaging. Ca2+metabol-ism disorders will cause cells reversible or irreversible damage, and then result in cellularstructure dysfunction. Mg2+is activator for a variety of intracellular enzyme, it can main-tain a stable internal structure of cells, and is conducive to normal growth and skeletalneuromuscular excitability. Abnormal Mg2+concentration will cause neurological damage.On the other hand, as the basic unit of the real life activities, individual cells (even thesame type of cells) is not exactly the same as each other. With the development of experi-mental techniques in recent years, more quantitative and objective evidences indicate thatin many critical life processes, such as embryonic development, cell differentiation, and thedevelopment and progression of the disease, the specific behavior of individual cells, aswell as the individual differences and heterogeneity, result in an extremely important oreven decisive results. Thus, in a single cell level and molecular level, accurately gras-pingthe content level of Na+, K+, Ca2+, Mg2+is the premise and basis of in-depth under-standing of their physiological and pathological mechanisms.However, due to Na+, K+, Ca2+, Mg2+not only co-exist and act synergistically in cells,but also similar in nature and interfere with each other, detection technology with high se-lectivity and efficient separation capacity is need; on the other hand, single cell is smalland intracellular component content is low with a large difference in concentration, so sin-gle cell analysis would require detection technology which is capable of handling a sampleof ultra-small size (single cell) and having a high detection sensitivity. The available resultsof biochemical analysis tools currently are mostly macroscopic observation or overall av-erage, and it is difficult to get the real information within such substances of single cell. Therefore, to reveal the relationship between Na+, K+, Ca2+, Mg2+and cell function regula-tion and disease, we must understand the true message of quantitative information within asingle cell of this material and the mutual synergy between them, and the development ofnew ways with highly sensitive, multi-parameter, and simultaneous quantitative determina-tion of metal ions (Na+, K+, Ca2+, Mg2+) within a single cell are in the urgent need.Based on this, in this paper, qualitative and quantitative analysis of various metal ions(Na+, K+, Ca2+, Mg2+) at the same time in single cell was explored and a new method thatcan be used for simultaneous detection of a variety of metal ions within a single cell wasdeveloped. Using this method, the content of metal ions within a single cell was determi-nated and the association of intracellular content and cell physiology and pathology waspreliminary inquiried. This thesis mainly includes the following four parts:The first chapter is preface, we first introduced briefly the biological function ofintracellular Na+, K+, Ca2+, Mg2+, and the current main detection methods and limitationsof intracellular metal ion, and then summed up the significance of single cell analysis andthe main research methods of components analysis of single cell, followed with detaileddescribing the composition and single-cell components analysis research progress with mi-crofluidic chip electrophoresis system. Finally the research status of the microfluidic sys-tem for intracellular metal ion detection was overviewed.In the second chapter, for single-cell Na+and K+synergies as well as the current dif-ficulties in the simultaneous quantitative detection, we developed a new methods in thedetection of Na+and K+at the same time in single cell based on cBDP fluorescent probeassociated with microchip electrophoresis. The fluorescent probe selectively responds toboth Na+and K+. The microfluidic chip electrophoresis achieves efficient separation of twocomponents, and high sensitive detection is obtained through laser-induced fluorescencedetector. The established method is simple, fast, and further applied to comparison of thesodium and potassium ion concent in normal cells and cancer cells and ion content changeduring the early apoptotic cells. The method is expected to provide a new way for the studyof biological problems related to sodium and potassium ions.In the third chapter, in view of single-cell Na+, K+, Ca2+, Mg2+physiological functionand interrelated effects, based on the second chapter, we developed a method to realized simultaneous quantitative detection of Na+, K+, Ca2+, Mg2+four ions in a single cell. Threefluorescent probes can specifically identify the four ions, and microfluidic chip electro-phoresis combined with multi-exciting and multi-emitting laser induced fluorescence de-tection platform enables effective separation and simultaneous quantitative detection offour kinds of metal ions fluorescence derivatives in a single cell. The determination in asingle4T1cell was completed. The established method could provides a new strategy forthe various components quantitative analysis within a single cell.The fourth chapter is summary and outlook, this paper presented two analytical meansof simultaneous multi-component determination within a single cell, and two methods areessentially the same and progressive. Firstly, single-cell Na+and K+simultaneous detectionwas achieved. Then on the basis of Na+and K+detection, we broadened the scope of ap-plication and developed a more applicable analytical method which is suitable for sin-gle-cell Na+, K+, Ca2+, Mg2+simultaneous determination. However, quantitative detectionat the same time for more intracellular metal ion and other moleculars related with cellphysiology and pathology also needs to be further discussed and studied. |
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