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Immunology experiment

Immunology experiment. Scores: 100 points Reports: 20% (principles, procedures, results, analysis of results ) Attendances: 5% Examination: 35% Experiment design: 40% Preparation: Isolation clothes, reagents Sanitation. Outlines. Humoral immunity function assay

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Immunology experiment

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  1. Immunology experiment • Scores:100 points • Reports: 20% (principles, procedures, results, analysis of results) • Attendances: 5% • Examination: 35% • Experiment design: 40% • Preparation:Isolation clothes, reagents • Sanitation

  2. Outlines • Humoral immunity function assay • Antibody preparation,detection • Detection Ag orAb in vitro • Cellular immunity function assay • Separation, proliferation of PBMC • Non-specific immunity function assay • Phagocytosis of macrophage • Immune pathology • Hypersensitivity

  3. Antibody preparation and detection ---- Ⅰ、Ⅱ、Ⅲ、Ⅳ p146 • Agglutination reaction ---- Ⅰ、Ⅱ p2 • Precipitation reaction ---- Ⅱ p6 • Antibody formation ablitiy ---- Ⅲ p139 • Complement fixation reaction ---- Ⅴ p7 • Immunological labeling technology ---- Ⅴ p10

  4. ExpⅠ Antibody preparation and Agglutination reaction (1)

  5. Test 1 Primary immunization

  6. Test 1 Primary immunization 1. Principles:

  7. Test 1 Primary immunization 1. Principles: • Xenoantigen: • Laws of Ab production mouse SRBC

  8. Test 1 Primary immunization 1. Principles: Ag Animals Serum(Contains Ab) Subcutaneously, 3 times

  9. Test 1 Primary immunization Uncoagulated SRBC 2. Methods: Add 3ml N.S and mix 2000rpm, 5min Discard the supernant Add 3ml N.S and mix 2000rpm, 5min Discard the supernant Take 0.1ml to immunize the mice subcutaneously Make 20%、25%、30%、40% SRBC solution

  10. Test 2 Agglutination reaction

  11. Hv Lv Lc Hc Review • Ag • Ab • Ag:Ab reaction

  12. Ⅰ Natureof Ag:Ab Reactions • Lock and Key Concept: high-specificity • Non-covalent Bonds • Hydrogen bonds、Electrostatic bonds、Van der Waal forces、Hydrophobic bonds • Reversible • Two phases: Binding phase Visible phase

  13. Ⅱ Factors Affecting Measurement of Ag/Ab Reactions • Affinity and Avidity • Ag:Ab ratio

  14. Ag: Ab ratio

  15. Agglutination reactions: paticulate Ag Precipitation reactions: soluble Ag Ⅱ Factors Affecting Measurement of Ag/Ab Reactions • Affinity and Avidity • Ag:Ab ratio • Physical forms of the antigen • Temperature: 37℃~56℃

  16. Effect of temperature

  17. Ⅱ Factors Affecting Measurement of Ag/Ab Reactions • Affinity and Avidity • Ag:Ab ratio • Physical forms of the antigen • Temperature: 37℃ • PH: 6~8 • Electrolytes: 0.9% NaCl

  18. Ⅲ Classical Ag-Ab reactions • Agglutination reaction • Precipitation reaction • Complement fixation reaction • Neutralization reaction

  19. Particulate Ag + Corresponding Ab Ⅳ Introduction of agglutination reaction • Definition: Visible agglomerate • Classification: Direct agglutination reaction: slide agglutination tube agglutination Indirect agglutination reaction

  20. Particulate Ag + Corresponding Ab Soluble Ag Visible agglomerate Direct agglutination reaction + Sensitized particulate Corresponding Ab Visible agglomerate Indirect agglutination reaction

  21. Test 1 Assay of blood type (ABO) • Principle: • ABO Blood type: • Slide agglutination reaction: • Material: • Anti-A serum and Anti-B serum • Slide • Needle • Woodstick Anti-A serum OR Anti-B serum ？ +

  22. Test 1 Assay of blood type (ABO) A B a. Slide • Method: Anti-A Anti-B Disinfect the finger b. Pick blood Stab the finger Drop one drop of blood into the serum c. Reaction Mix blood with the serum by woodstick d. observation

  23. Test 1 Assay of blood type (ABO) Blood type • Anticipated results: A B AB O

  24. Test 1 Assay of blood type (ABO) • Attentions: • Keep the serum seperated • Disinfection • Do not confuse the two sides of wooden stick • Observe the result in time • Put the blood-stained material in a certain place

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