Used In The Study And Application Of In Situ Imaging Of The Metal Ions In Living Cells

With the progress of biochemistry and molecular biology and the using of chemistry, physics and biology, the course of chemical reaction in organism was elucidated gradually. But more and more experimental evidences suggest that the results obtained from the reaction occurred in test tubes cannot reflect the instance in living object. So it is necessary to explore the real time changes of active substances in order to comprehend the life.Fluorescence analysis and imaging techenology was widely used in biological analysis field because of their high sensitivity and selectivity. Most of biological cells have no or weak fluorescence, which make their sensitivity very low. The interference was lowed largely. Fluorescent probe was widely used in biological science field because of their special photophysics and photochemical characters, high sensitivity, wide response range and appropriate condition. So we need to design and synthesize fluorescence probe with appropriate spectrum character. Then we explore the property in chemical circumstance and by integrating with fluorescence imaging technology to detect the concentration changes of metal ions and the distribution in cell on line. We then explored the function of metal ions in life activity and pathology of the shortness of metal ions to achieve the detection of reactive oxygen species in vivo and in situ, explore the pathology of reactive oxygen species burst, which will afford essential basis for biology and medicine study.Trace metal ions in organisms play essential roles in many biological processes. Among them, redox-active copper ion is a crucial element in enzyme functions and transcriptional events. An alteration of intracellular copper ion homeostasis is connected to various neurodegenerative diseases, including Menkes and Wilson diseases, familial amyotropic lateral sclerosis, Alzheimer's disease, and prion disease.Thus, on one hand, appropriate level of Cu2+ is important for life; on the other hand, excessive Cu2+ is highly toxic to organisms. For these reasons, the quantitative detection of intracellular Cu2+ is of great importance for elucidating the complex physiological and pathological roles of copper.Protons are one of the most important targets among the intracellular species of interest, as it is well-known that intracellular pH plays a central role in many cellular events such as cell growth, calcium regulation, endocytosis, chemotaxis, cell adhesion, and other cellular processes. Thus, it would be extremely desirable to have access to a broad variety of"acidic"probes with high sensitivity and selectivity, good photostability and membrane-permeable.In addition, anions present in the human body, play essential roles in many biological processes. Particularly, fluoride is indispensable trace element in creature vital activity. The effect of fluoride on humans has a dual role as an essential element and at high levels as a toxic substance. Excess fluoride ion results in fluorosis and renal, gastrointestinal and immunological toxicity. Thus, the process of quantifying trace fluoride anion within biological samples, elucidation of the complex physiological and pathological roles provide motivation for developing new ways to study dynamic fluoride chemistry in living systems. Up to now, there is no small molecules have ever been available for detection and imaging fluoride in living biological systems.Based on the changes in spectrum characters of the fluorescent probes reacting with metal ions, protons and fluoride, we have carried out three aspects of investigation: First, we designed and synthesized a novel fluorescence probe based on rhodamine dyes for Cu2+ and optimized the detecting condition. The reaction mechanism was studied and the proposed methods were then applied to determine Cu2+ in RAW264.7 MacrophagesSecond, designed and synthesized an acidic fluorescent probe based on rhodamine dyes for H+ and optimized the detecting condition. The reaction mechanism was studied and the proposed methods were then applied to determine H+ in HepG2 Cells Third, these papers presented the synthesis a new fluorescent probe and assume the probe can detect fluoride in aqueous solutions.