Isolation of chloroplast andfluorescence observation.pptVIP

Isolation of chloroplast and fluorescence observation 叶绿体分离和荧光观察 Instructor ：Feng Lijun， Ph.D. 授课老师：冯力骏 drfeng@sdu.edu.cn The structure and location of Chloroplast Chloroplasts are specialized organelles containing chlorophyll, which provides the green color. They are one of the two double membrane organelles in the cell. Within the stroma are the thylakoids and grana where photosynthesis takes place. 叶绿体内含叶绿素，使植物呈现绿色。 叶绿体是细胞内三个双层膜细胞器之一。 叶绿体内基粒是光合作用的场所。 Particles’ sedimentation rate in a centrifugal field depends on the particle’s size, shape and density, also the centrifugal force and the viscosity of the suspension medium. By control the centrifugal force and centrifugation time in proper order, different organelles could be separated. 在离心场里，颗粒的沉降速度取决于颗粒的大小、形状和密度，还有离心力和离心介质的粘度等因素。 通过控制离心力和离心时间等因素，不同的细胞器可以通过阶梯离心的方式得到分离。 Light energy absorbed by chlorophyll is mainly used to drive photosynthesis. Excess energy can be dissipated as heat or can be re-emitted as light—chlorophyll fluorescence. 叶绿素吸收的光能主要用于光合作用。 多余的能量可能以热的形式散发，也可能以激发光的形式散发。 这种激发光就称为叶绿素荧光。 Schematic Diagram of the Fluorescence Microscope. Chlorophyll fluorescence analysis has become one of the most powerful and widely used techniques available to plant physiologists and ecophysiologists. 叶绿素荧光检测技术作为一种强大的工具，广泛应用于生态学和植物生理学研究。 Purpose of this experiment本次实验目的 Learn to isolate chloroplasts through serial centrifugation. 利用梯度离心技术分离叶绿体。 Observe fluoresence of chloroplasts. 观察叶绿素激发荧光。 Detailed protocol description and lab notes 学习试验过程的详细描述及实验记录 Learn scientific writing and presentation 学习科研报告的写作和展示 Materials 材料 Reagents: 0.35M NaCl solution fresh spinach Tools and equipments: homogenizer, balance, centrifuge, beaker, cloth, conventional light microscope, fluorescence microscope, coverslip, glass slide, pipette, forceps. 试剂： 0.35M氯化钠等渗溶液 新鲜菠菜 仪器、工具： 天平，匀浆机，离心机，烧杯，纱布，普通光学显微镜，荧光显微镜等。 Methods： Weigh 3X30g fresh spinach leaves , removing stems and thick veins.