The research on phase-based lifetime detection of zebrafish interior region the blood oxygen content distribution sensing

• Main Authors: Yu, Chu-Hung, 游礎鴻
• Other Authors: Huang, Shih-Hao
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/52162299055328286120

碩士 === 國立臺灣海洋大學 === 機械與機電工程學系 === 102 === This study adopted the microfluidic chip integrated with the phosphorescence phase difference optical detecting system to develop the device for detecting local distribution of blood and oxygen in the body of juvenile fish of single zebra fish. The microfluidic detecting platform combined with two components: the pneumatic actuator positioning zebra fish microfluidic chip and the phosphorescence phase difference optical inspection system. The pneumatic actuator positioning zebra fish microfluidic chip was used to localize zebra fish in the flow channel to culture for a long time, and to change external environment by changing injected fluid, or to immerse fish in agents for a long time, and to carry off effluent via peristaltic pump, thus maintaining a stable stimulation of agents and environment. Role of pneumatic actuator positioning zebra fish device: pneumatic deformation of upper flow channel allowed fish to be localized only in wider flow channel than its body width. Each organ or tissue can be measured for a long time under the stable localization. Phosphorescence phase difference optical inspection system was used to control the light source projection figure of Blue-Laser through digital micromirror device (DMD) within projector and to stimulate blood oxygen value in blood in the area for detection, and then achieved the automatic real-time effects of stimulated measurement.we used Oxyphors G4, Pd-meso-tetra-(3,5-dicarboxyphenyl(referred as Oxyphors G4 below) as the phosphorescence sensing measurement index within juvenile fish of zebra fish. The index was injected into fish bodies which were grown-up for 48h. In comparison of common phosphate buffered solution (PBS) to HEPES as hydrogen ion concentration buffer in water flea injection, HEPES can maintain the PH in 7.2-7.4 which is near the PH value within fish body for a long time to lowering motality. 9.2nl of HEPES with lower lethality was injected in a concentration of 200uM to establish the sensing system in vivo. The experimental findings founded that : the ratio of blood and oxygen in fish body was decreased gradually to 0% within 10 minutes in the low oxygen environment (0.01 mg/L O2); however, the ratio of blood and oxygen of some fishes were recovered to the ratio within the general water body under the recovered oxygen environment(7-8 mg/L O2). We measured the changes in blood and oxygen within two different blood pooling areas in zebra fish body and detected the difference of the blood and oxygen variability in external extreme environment between the loop valley and caudal veins, and the initial value of ratio of blood and oxygen in veins was lower, also the blood oxygen level was decreased quickly to 0%. This study can establish a measurement way for the detection mechanism in the body of living organisms. It is expected that this blood oxygen sensing system can be applied to the micro-tumors model of animals like zebra fish in future to observe the correlation between active state of cancer cells to the blood and oxygen