Immunohistochemistry
Imunohistoquímica
Course code:68310
José Ferreira da Silva
Learning outcomes of the curricular unit:
INSTRUMENTAL COMPETENCE
To complete the frequency of this discipline the student should:
- Understand the historical contextualization of the imunocytochemistry techniques and its current importance in pathological diagnosis;
- Define and characterize enzymes, taking into account their structural and functional features; Understand the processes involved in enzymatic specificity in relation to substrates;
- Understand the influence of some factors on enzyme catalysis;
- Understanding the process underlying the classification of enzymes;
- Define and characterize antibody and antigen taking into account structural and functional characteristics;
- Describe how antibody and antigen cooperate on ICQ for the formation of antigen-antibody complex;
- Know the characteristics of some panels of antibodies;
- Understand the importance of quality control in ICQ.
SYSTEMIC COMPETENCE
- to identify and characterize most commonly used enzymes in ICQ
- Identify, interpret and criticize the various methods and techniques in ICQ.
- Recognize and relate the various factors that influence the methods and techniques ICQ;
- Planning techniques and methods specific diagnostic situations ICQ and research;
Syllabus:
- Presentation the program and evaluation form of theoretical and practical classes; Introduction to ICQ techniques; Historical facts of immunocytochemistry techniques; Usefulness of ICQ.
- Enzymology: historical facts; characterization of enzymes; enzymatic specificity; classification of enzymes;
- Enzymology: factors influencing enzyme kinetics, such as temperature and pH, among others. Presentation and characterization of some enzymes used in the context of immunocytochemistry techniques.
- Antibody: synonyms; structure, function and classification of immunoglobulins; Definition, characterization and classification of antigens. Characterization of the antibody-Antigen complex. Rules of handling of antibodies in the lab environment.
- Methods for Immunolabeling and its characterization: Immunofluorescence; Imunoouro and techniques of enzymology. Classification of methods according to number steps: direct methods; indirect methods; Classification of immunoenzymatic methods: PAP (Peroxidase anti peroxidase); APAAP (anti alkaline phosphatase alkaline phosphatase) and avidin-biotin methods (ABC-avidin biotin complex and LSAB-Labeled Streptavidin-Biotin).
- Revisions
- Diagnostic assessment – do not account for note
- Methods of direct and indirect polymer. Other methods such as ImpressTM and CSATM. Advantages and disadvantages of immunoenzymatic methods. Double immunolabeling sequential and simultaneously. Characterization of molecules of streptavidin, biotin and avidin.
- Presentation and characterization of substrates and chromogens more used. Equipment used in the incubation times of serum.
- Causes and approaches to prevent the nonspecific immune labelling
- Production of monoclonal and polyclonal antibodies: characterization and production technologies
- Fixation and effect on the antigen. Reversal effect of ion calcium complexion and protein cross-links formed by formalin fixation (antigen retrieval techniques in immunohistochemistry)
- ICQ applications: primary antibodies for leucocitary antigens, intermediate filaments antigens and epithelial antigens.
- ICQ Applications: primary antibodies for cellular proliferation, hormone receptors; oncoproteíns; other primary antibodies.
- Quality control in ICQ.
- Reviews
Suggested Bibliography:
- Boenisch T et al (2001) Handbook of immunochemical staining methods. 3rd ed., Dako Corporation.
- Dabbs DJ (2018) Diagnostic Immunohistochemistry. 5th ed., Elsevier.
- Suvarna K, Layton C, Bancroft J (2018) Theory and Pratice of Histological Techniques. 8th ed., Elsevier.
- Polak J, Noorden SV (1997) Introduction to immunocytochemistry – BIOS Scientific Publishers.
- Polak J; Noorden SV (1986) Immunocytochemistry - Modern methods and applications. 2nd ed., Wright