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Immunocytochemical methods and protocols / edited by Constance Oliver and Maria Célia Jamur. — 3rd ed. — New York : Springer, c2010. – (58.17/M592/v.588) |
Contents
Contents
Contributors
PART I ANTIBODY PREPARATION
1. Overview of Antibodies for Immunochemistry
2. Introduction to the Purification of Antibodies
3. Antibody Purification: Ammonium Sulfate Fractionation or Gel Filtration
4 Antibody Purification: Ion-Exchange Chromatography
5. Antibody Purification: Affinity Chromatography-Protein A and Protein G Sepharose
6. Conjugation of Fluorochromes to Antibodies
7. Biotinylation of Antibodies
PART II TISSUE PREPARATION FOR LIGHT MICROSCOPIC ANALYSIS
8. Cell Fixatives for Immunostaining
9. Permeabilization of Cell Membranes
10. Preparation of Frozen Sections for Analysis
11. Processing of Cytological Specimens
12. Processing of Tissue Culture Cells..
13. Processing of Tissue Specimens
14. Heat Induced Antigen Retrieval for Immunohistochemical Reactions in Routinely Processed Paraffin Sections
PART III LIGHT MICROSCOPIC DETECTION SYSTEMS
15. Fluorochromes-Properties and Characteristics
16. Direct Immunofluorescent Labeling of Cells
17. Fluorescence Labeling of Surface Antigens of Attached or Suspended Tissue-Culture Cells
18. Fluorescence Labeling of Intracellular Antigens of Attached or Suspended Tissue-Culture Cells
19. Fluorescent Visualization of Macromolecules in Drosophila Whole Mounts 168
20. Overview of Conventional Fluorescence Photomicrography
21. Overview of Confocal Microscopy
22. Overview of Laser Microbeam Applications as Related to Antibody Targeting
23. Immuno-Laser Capture Microdissection of Rat Brain Neurons for Real Time Quantitative PCR 219
24. Overview of Antigen Detection Through Enzymatic Activity
25. The Peroxidase-Antiperoxidase (PAP) Method and Other All-Immunologic Detection Methods
26. The Avidin-Biotin Complex (ABC) Method and Other Avidin-Biotin Binding Methods
27. Avidin-Biotin Labeling of Cellular Antigens in Cryostat-Sectioned Tissue
28. Multiple Antigen Immunostaining Procedures
29. Immunoenzymatic Quantitative Analysis of Antigens Expressed on the Cell Surface (Cell-ELISA) 301
30. Use of Immunogold with Silver Enhancement 311
PART IV FLUORESCENCE-ACTIVATED CELL SORTER (FACS) ANALYSES
31. Overview of Flow Cytometry and Fluorescent Probes for Flow Cytometry 319
32. Tissue Disaggregation 327
33. Indirect Immunofluorescent Labeling of Viable Cells 331
34. Indirect Immunofluorescent Labeling of Fixed Cells 335
35. Fluorescent Labeling of DNA
36. Deparaffinization and Processing of Pathologic Material 345
PART V COLLOIDAL GOLD DETECTION SYSTEMS FOR ELECTRON MICROSCOPIC ANALYSIS
37. Fixation and Embedding
38. Preparation of Colloidal Gold 363
39. Conjugation of Colloidal Gold to Proteins 369
40. Colloidal Gold/Streptavidin Methods
41. Pre-embedding Labeling Methods
42. Postembedding Labeling Methods
PART VI THE CLINICAL LABORATORY
43. The Clinical Immunohistochemistry Laboratory: Regulations and Troubleshooting Guidelines 399
Index 413
