Preparation And Research Of Two-photon Ca²⁺ Fluorescent Probe

Ca²⁺acts as a universal messenger in a variety of cells. Numerous functions of alltypes of cells are regulated by calcium to a greater or lesser degree. Serving as anintracellular second messenger and mediator，Ca²⁺plays a very important role inintraeellular signal transduetion as well as extraeellular. Consequently， themeasurement of cytosolic Ca²⁺has become an important investigation area inchemieal，biological and medical researeh, and it is also successful applied on organicsynthetic chemistry.Fluorescence probes, provide a new method in the research of intracellular calcium.But the traditional fluorescent probe for is to use single pHoton to inspire, So therewould be a lot of not easy to solve problems, including ultraviolet light transmission sexis low, and the organization of the existence of living cell damage, can make the cellsthemselves a fluorescence effect observation will also have the light bleachingpHenomena exists, etc. But using two-pHoton as excitation wavelength of fluorescentprobe can be very good to avoid the negative effect, it is to use the infrared or nearinfrared to stimulate can better protect cells to avoid the damage to cells, still can go outof the cell fluorescence effect, and has good penetrating. Additionally, two-pHotontechnich can improve the spatial resolution and solve the deep imaging in biologicaltissue. Two-pHoton fluorescence probe has a trend to be used for biological research ofsmall molecules probeIn view of the advantages of two-pHoton fluorescence probe and thedisadvantages of traditional calcium ions probe, a new two-pHoton fluorescent Ca²⁺probe is designd and synthesized in this paper. A series of optical characterization arecarried out for it, and study the optical properties after the interaction with Ca²⁺.Themain contents of this paper is as follows:1、nthetic a two-pHoton fluorescent Ca²⁺probe containing the structure ofBAPTA, and a polymer of strong fluorescence properties. Vilsmeier and wittigreaction are the main use in the Synthesis of the probe.1HNMR, IR are utilized forcharacterization all final compounds.2、We studied the UV absorption spectra and one-pHoton fluorescence specta ofthe compound in different solvents （including water, ethanol, acetonitrile, DMF andacetone） and the different metal ions titrating; The UV absorption spectrum and one-pHoton fluorescence spectrum of the compound was titrated with the Ca²⁺ofdifferent concentrations; We studied the two-pHoton fluorescence spectrum of thecompounds interacting with the Ca²⁺of different concentrations at800nm.3、We studied the UV absorption spectrum、one-pHoton and two-pHotonfluorescence spectrum of the compound was titrated with the fluorescent polymer,meanwhile determined the best polymer using concentration under differentconditions...