| The Laser tweezers Raman Spectroscopy (LTRS) is the new technology that is a combination of Optical tweezers technology and Laser micro Raman technology to make the laser beam highly focused on the three dimensional space to trap particles and acquire its Raman spectrocopy using the high numerical aperture objective. LTRS technology has been mainly used to study single living suspension cells, microorganism and biological macromolecules. Since it has simple, quick, non-invasive, noninvasive, repeatable and high sensitivity advantages, it has become the most popular tool in researching single cells molecular structure.Blood glucose is the sugar in the blood, which is the source of energy needed for human life activities. Hypoglycemia can suffer from symptoms such as memory loss, unresponsive, dizzy fan; hyperglycemia can cause the lesions in the retina, myocardial and nerve tissue microvasculature. Keeping the normal blood glucose levels in the body is very important. Human body tissue cells are intake from the blood glucose, especially the red blood cells, mature red blood cells are without mitochondria and the nucleus, and the energy used for their own survival and metabolism is from glucose in the blood, so researching the blood glucose levels and the way of red blood cells to take in glucose is of great significance to keep blood sugar balance and the storage of red blood cells in the freeze drying way.Nitroglycerin has already one hundred years of history as a special effects commonly used drugs currently in prevention and treatment of angina pectoris. It took action fast, short duration and can quickly relieve angina, but its mechanism is still controversial in the body metabolism Nitroglycerin generated nitrite in the body has been reported earlier, it is generally recognized as inactive metabolites, produced by theoxidation of Nitric Oxide, which is with vascular regulating activity. Nitric Oxide acts on smooth muscle cells, make smooth muscle relaxation, cause vasodilation and blood pressure drops. Nitroglycerin in vivo metabolism involves a variety of metabolic enzyme activity, endogenous active substances level and the change of receptor activity, its metabolic pathway is not yet clear, studying the metabolic mechanism of nitroglycerin is of great significance.In this thesis, the LTRS was used to detect and analyze noninvasive blood glucose, explore the way of red blood cells to take in glucose. And we also applied LTRS to nude mice ear artery blood vessel noninvasive spectral analysis, taking the spectrum of blood after injection of nitroglycerin, paying. attention to the change of blood components and effects of time after injected nitroglycerin. Injection again after half an hour, collected Raman spectra and analyzed the extent of vasodilation, investigated the drug resistance of nitroglycerin.The first chapter introduced the background of Raman spectrum, the basic principle and two important parameters:Raman shift and Raman intensity. Secondly introduced the Raman spectra of some main characteristics, such as the Raman scattering intensity has a linear relation with the sample. Raman Shift reflected the molecular information of vibration and rotation, and had nothing to do with the excitation wavelength of the laser, samples were without preparation, contact and trauma, Aqueous solution was suitable for study the characteristics of biological samples. Then, this paper introduced the composition of Raman spectrometer:light source, optical path system, the chromatic dispersion system, the signal detection and processing system; Finally introduced the other Raman spectroscopy, such as Resonance Raman Spectroscopy, Surface Enhanced Raman Scattering, Fourier Transform Near-Infrared Raman Spectroscopy.Chapter two mainly introduced the LTRS. At first, we introduced the productive background of Optical Tweezers and its the basic principle and the application; Secondly introduced the LTRS system device, included Optical Tweezers system and Raman system, the main devices were semiconductor laser, spectrometer, microscope, CCD spectrometer, photoelectric coupling detector and some optical device. Finally this paper introduced the methods of Raman spectrum signal processing:baseline correction and normalization method.The third chapter used the LTRS noninvasive detection of blood glucose content, explored the way of red blood cells to take in glucose. The results showed that the Raman spectra of blood and red blood cells were the same, and mainly composited by the hemoglobin peaks of752,1001,1214,1450,1549,1618cm-1and glucose peak of1125cm-1. Plasma Raman spectra contained weak hemoglobin peaks, because heparin anticoagulant could keep blood no frezen and make retention for clotting factor and fibrin. Serum was a pale yellow liquid after the blood clot a dilution, clotting factor and fibrin was consumed in the process of condensation, therefore, serum didnot contain fiber protein, but contained trace amounts of fibrin in plasma. Red blood cells of1125cm-1peak was significantly higher than the plasma. Making standard glucose solution concentration curse was to measure the concentration of glucose in red blood cells and plasma, Anthracene copper colorimetric method showed that blood glucose levels in red blood cells was about1.5times that of the plasma. Using different concentrations of glucose solution to treat the blood, found that with the increasing of glucose concentration, glucose with the help of some special membrane protein in membrane structure could rapidly into the red blood cells. Red blood cells took in glucose not only by facilitated diffusion, there would be an active way of transportation.In Chapter four, Raman spectra of blood in arteries of nude mice injected with nitroglycerin were analyzed in real-time and noninvasively. The Raman spectrum of Blood in artery was captured every10seconds. Analysis results show that the spectra of the blood mainly have these characteristic peaks:1548,1618,1654cm-1(protein) and1125cm-1(blood sugar). Raman spectral intensity decreased from280to730seconds after the injection of nitroglycerin. This may be due to the expansion of blood vessels with nitroglycerin. The characteristic peak intensities before and after nitroglycerin injection changed as below:the1548cm-1peak intensity decreased from1965.42to1273.61by35.2%; the peak1125cm-1intensity decreased from411.59to223.79by46.63%. This means that the content of protein and sugar in blood had decreased, namely, hemoglobin in the blood of nude mice had denatured. To study the tolerance of nitroglycerin, at the first injection of nitroglycerin and then injected the same amount of nitroglycerin again after half an hour, The results showed hemoglobin characteristic peaks at second drop degree were inferior to the first time, and consumed more reaction time, and nude mice after continuous injection of nitroglycerin have had drug resistance. The reason for this may be because of nitroglycerin generate NO need to consume hydrophobic base (SH) and cysteine, and when injected nitroglycerin at the second times, cysteine couldn’t be added in time, NO generation decreased, so the effect of nitroglycerin weakened. |
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