《分子細胞生物學實驗教程》為外招生專業(yè)教材,全英文寫作,是關于分子細胞生物學實驗的一本實驗操作指南。正文部分分為十一個章節(jié),包含從兩個層次對分子細胞生物學當中的細胞分離、DNA分離與純化、蛋白質的制備電泳、細胞遺傳等內容,適合從事相關研究工作的人員參考閱讀。
《分子細胞生物學實驗教程》由曁南大學出版社出版。
Author's Comments
Chapter 1 Microscopy
1.1 Introduction: Light Microscope--Resolution vs Magnification
1.2 Bright Field, Dark Field, Phase Contrast
1.3 Fluorescence Microscopy and Confocal Microscopy
1.4 Electron Microscopy
1.5 Exercise : Use of a Bright Field Microscope
1.6 Exercise : Examination of Different Cell Types
Chapter 2 Subcellular Structures
2.1 Introduction: CeBs, Organelles and Vital Staining
2.2 Exercise : Vacuole in Animal Cells
2.3 Exercise : Staining of Mitochandria in Human Cheek Epithelial Cells
2.4 Immuno-Staining and Fluorescent Labeling
2.4.1 Immuno-Staining
2.4.2 Fluorophores and Fluorescent Labeling
2.5 Exercise: Inmunofluorcscent Analysis of Microtubules
Chapter 3 Centrifugation and Subcellular Fractionation
3.1 Introduction: Equilibrium Density Gradient Centrifugation and Differential Sedimentation
3.2 Exercise: Purification of Nuclear and Mitochondria by Differentia Sedimentation
3.3 Exercise: Assay of Fractions for Succinate Dehydrogenase (SDH)
Chapter 4 Cell Membrane and Cross-Membrane Transport
4.1 Introduction: Membranes and Its Properties
4.2 Exercise: Osmosis
4.3 Exercise: Hemolysis
4.4 Exercise : Pinocytosis
Chapter 5 Cell Cycle and Cell Division
5.1 Introduction: Types of Cell Division
5.2 Exercise: Study of Different Stages of Mitosis in Onion Root Tip Cells
5.3 Exercise : Study of Different Stages of Meiosis in Mouse Testis Cells
Chapter 6 Cytogeics and Karyotyping
6.1 Introduction: Karyotyping
6.2 Exercise: Short Term Culture of White Blood Cells and Preparation of Metaphase Chromosomes
6.3 G-and C-Banding of Metaphase Chromosomes
6.3.1 Exercise : G-Banding of Metaphase Chromosomes
6.3.2 Exercise : C-Banding of Metaphase Chromosomes
6.4 Exercise : Karyotyping
Chapter 7 Cell Culture
7.1 Introduction: Principles of Animal Cell Culture
7.2 Sterilization Techniques
7.3 Exercise : Establishment of a Primary Culture
7.4 Exercise: Cell Viability Assay by Trypan Blue Exclusion
7.5 Exercise: MTT Assay for Cell Viability
7.6 Exercise : Mammalian Cell Transfectian
7.6.1 Exercise : Lioofectamine Mediated Mammalian Cell Transfection
7.6.2 Exercise: Transfecting Mammalian Cells by Electroporation
7.7 Exercise: PEG Induced Fusion of Erythrocytes
Chapter 8 Apoptosis Analysis
8.1 Introduction : The Characteristic Cellular Changes of Apoptosis
8.2 Exercise: Induction of Apoptosis
8.3 Exercise: Vital Staining and Detection of Chromatin Condensation
8.4 Exercise : Detection of DNA Fragmentation
Chapter 9 Flow Cytometry
9.1 Introduction: Flow Cytometry and Signal Processing
9.2 Exercise: Cell Cycle Analysis by Propidium Iodide (PI) Staining
9.3 Exercise: Cell Sorting by Fluorescence-Activated Cell Sorting (FACS)
Chapter 10 Cellular Immunology: Enzyme-Linked Immunosorbent Assay (ELISA)
10.1 Introduction: The History and the Application
10.2 Exercise: Direct ELISA (Also Known as Sandwich ELISA)
10.3 Exercise: Capture ELISA
Chapter 11 Molecular Cell Biology
11.1 Exercise: DNA Extraction and Purification
11.2 Exercise : Determination of DNA Concentration Using the Spectrophotometer
11.3 Exercise : Total RNA Isolation from Cells in Culture
11.4 Standard PCR
11.4.1 Introduction : Principles of PCR
11.4.2 Exercise : PCR Amplification of DNA
11.4.3 Exercise: Two-Step RT-PCR Amplification of B-Actin
11.5 Exercise: Southern Blot Using 32p Labeled d GTP
11.6 Measurement of Call Protein Content
11.6.1 Introduction: Bradford Assay
11.6.2 Exercise : Protein Assay and Measurement of Protein Content of Cells
11.7 Protein Extraction and Purification
11.7.1 Introduction : The Principles for Protein Extraction
11.7.2 Exercise : Whole Cell Extract Preparation
11.8 Exercise : Protein Electrophoresis
11.8.1 Introduction: Protein Electrophoresis and SDS-PAGE
11.8.2 Exercise : Protein Separation on SDS-PAGE
11.9 Protein Analysis and Detection
11.9.1 Exercise : Visualization of Proteins in Gels
11.9.2 Exercise : Semi-Dry Transfer of Protein onto Membrane
11.9.3 Exercise: Western Blot
Appendix : Chemical Preparations
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Two types of electron microscopy-transmission and scanning-are widely used to study cells.The transmission electron microscope (TEM) has a resolving power of 3A-10A. The scanning electron microscope (SEM) is being increasingly popular with cell biologists because of its remarkable ability for quantifiable mapping of surface detail, along with improved resolution (30A-iooA) and its ability to show 3D structure.
The transmission electron microscope is identical in concept to the modern light microscope. It is posed of a light source (in this case an electron source), a substage condenser to focus the electrons on the specimen, and an objective and ocular lens system. In the electron microscope, the ocular lens is replaced with a projection lens, since it projects an image onto a fluorescent screen or aphotographic plate.Since the electrons do not pass through glass, they are focused by electromagic fields. Instead of rotating a nose-piece with different fixed lenses, the EM merely changes the current and voltage applied to the electromagic lenses.
Another characteristic of electron microscopes is that they are usually designed upside-down, similar to an inverted light microscope.The electron source is on top, and the electrons travel down the tube, opposite to light rays traveling up a microscope tube.This is merely a design feature that allows the operator and technicians ease of access to its various ponents. The newer electron microscope is beginning to look like a desk with a TV monitor on it.