细胞生物学英文课件：细胞生物学研究方法.pptVIP

六. Structure determination of biomacromolecule nuclear magnetic resonance spectroscopy（NMR ,核磁共振光谱仪) X-ray diffraction (X-射线衍射技术) 七. Cell engineering （一） Techniques in cell engineering 1. Bulk culture (大规模培养) Suspension culture Immobilization culture Microcarriers culture Three-dimensional cell culture 2. Cell fusion (细胞融合) 3. Nuclear transfer (细胞核移植) 4. Gene transfer (基因转移) （二） Application of cell engineering 1. Monoclonal antibody 2. Pharmaceutical protein 3. Cell therapy Characterization Cell proliferation (BrdU, PCNA) Cell viability/death (Live/Dead staining) Cell differentiation - Neurogenesis Neuronal generation (TuJ1, MAP-2, RT-PCR) Ion channels/receptors (Ca2+ imagimg) Synaptic activity (FM1-43) - Gliogenesis Glial generation (GFAP, O4, RT-PCR) Transplantation Mechanical dissociation Stem cells LV III IV SC cc Rat Embryos Cell-Collagen Constructs Type I Collagen Cell-Hydrogel Constructs Proliferative Neural Stem and Progenitor Cells Differentiated Neuronal Networks Brain-like Tissues Synthetic Nanofibers 100mm Rotary Wall Vessel Bioreactor B GFAP(red)/BrdU(green) Control Transplanted 三、Isolation and purification of cellular component 1. Differential centrifugation (差速离心法) 2. Velocity sedimentation (速度沉降) 3. Equilibrium sedimentation（平衡沉降） 4. Column chromatography (柱层析) 5. SDS(SDS聚丙烯酰胺凝胶电泳) 6. Isoelectric focusing (等点聚集电泳) 7. Two-dimensional electrophoresis (双向电泳) 8. Isolation, purification and identification of nuclear acid Differential centrifugation (差速离心法) Electrophoresis (电泳) Agarose gel electrophoresis picture 四、Cytochemistry and labeling in alive cells 1. Enzyme cytochemistry (酶细胞化学技术) 2. Immunocytochemistry (免疫细胞化学技术) 一抗：rb, mouse 二抗：羊抗兔，羊抗鼠 用兔子IgG去免疫绵羊 标记荧光素 (一) Cytochemistry 3. Autoradiography (放射自显影技术) 放射性标记的有丝分裂标记法（percentage labeled mitoses，PLM）：原理是对测定细胞进行脉冲标记、定时取材、利用放射自显影技术显示标记细胞，通过统计标记有丝分裂细胞百分数(PLM)的办法来测定细胞周期. (二) Labeling in alive cells 用钙离子探针观察细胞内钙离子的动态变化 1. Ion microprobe 2. Gre