The Application Of Chemiluminescence Probe In Superficial Tumour Diagnosis And ROS Detection

Reactive oxygen species (ROS) is produced continuously in biology tissue. The quantities of some ROS are few, while the others are great and can be detected by some detective methods. With the extension and penetration of the life science research, people have gradually realized that the occurrence and deterioration of many kinds of diseases and the degradation of organism functions have close relationship with the activities of ROS, such as atherosclerosis, neuro-degenerative diseases, carcinogenesis. Accordingly, detection the generation, distribution and content of ROS inside the biologic system have great significance to fundamental and clinical researches.In this paper, the research work is that using Cypridina luciferin analog (CLA) as a sensitive and selective CL probe to apply the high sensitivity optical imaging detection technology to the measurement of ROS generated in biological system. Furthermore, a novel detection method for singlet and superoxide anion is reported in paper and the mechanism is study initially. In this paper, the generation path and detection methods of singlet are introduced firstly, then the characteristic of Cypridina Luciferin Analog is described in detail.The first experimental study mainly include that an in vivo usable chemiluminescence probe carried by nanoparticles and its application in Photodynamic Diagnosis (FDD). The chemiluminescence probe is to coat MCLA with albumin. This can decrease loss of MCLA in transportation process in tissue. Electron microscope observation indicated that the diameter of nanoparticles was about 50~150nm. With blood circulation, nanoparticles can uniform distribution finally in organism. The results in vitro and in vivo showed that the nanoparticles could release MCLA slowly. In vivo experiment, injected the Hematoporphyrin Derivative (HpD) and nanoparticles into a mice carried tumor via tail vein, stimulated by light, then can see distinct luminescence at the site of tumor through detecting theinABSTRACTtwo-dimensional image using a highly sensitive Intensified Charged-Coupled Device (ICCD) detector. The results of experiment proved that the time delay resulted from nanoparticles is obvious.In the second experimental study, a novel detection method for singlet and superoxide anion is presented. This method can be used to selectively detect 'Oi and C>2~ with high-specificity and high-sensitivity. With the single photon counting system, very weak chemiluminescence could be detected from the FCLA solution at natural partial oxygen pressure. When human serum albumin was added into the FCLA solution, a strong enhancement of the chemiluminescence with a peak at 525nm was observed. When reducing the concentration of oxygen in the solution by blowing with Na until oxygen approaches zero, chemiluminescence almost disappeared. These experimental phenomena proved that the oxygen molecules dissolved in the solution play a very important role to the chemiluminescence contributions. Based on these facts, a novel high sensitive detection method of '(^ and 02 ~ was proposed. The principle of this method is similar to that for a measurement of ATP using luciferin and luciferase. The method has high sensitivity and high specificity, so it will be have enormous application potency in fundamental research and clinical medicine.