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14D2I08 - Methods of examination of cells, tissues and organs in physiological and pathophysiological conditions

Course specification
Course title Methods of examination of cells, tissues and organs in physiological and pathophysiological conditions
Acronym 14D2I08
Study programme DOCTORAL ACADEMIC STUDIES
Module
Type of study PhD academic studies
Lecturer (for classes)
Lecturer/Associate (for practice)
    Lecturer/Associate (for OTC)
      ESPB 10.0 Status elective
      Condition Enrolled semester Oblik uslovljenosti
      The goal The aim of the course is to train the students for basic procedures in the laboratory work, for collection and preservation of samples and to obtain basic theoretical and practical knowledge of various methods that can be used to investigate the structure and function of cells, tissues and organs in the physiological and pathophysiological conditions.
      The outcome The independence of the student in the collection and processing of samples, as well as the possibility of student to combine different laboratory methods in research.
      Contents
      Contents of lectures Good laboratory practice, laboratory dishes, washing, sterilization. Making solutions, making the standard curve. Blood sampling, anticoagulants, short-term and long-term storage of samples. Sampling of various body fluids. Methods of sampling tissue for biochemical, immunohistochemical and histological analysis. Methods for sampling various organs/tissues for biochemical and toxicological tests. Staining of blood smears and cells from body fluids. Histological techniques. Protein concentration, isolation and purification of proteins, protein separation methods : native and SDS PAGE electrophoresis, chromatography. Specifique protein determination and quantification : ELISA , RIA, Western blot, immunodiffusion, immunohistochemistry, immunocytochemistry. Flow cytometry. Determination of the activity of the enzyme and isoenzymes: spectrophotometry, cytochemical, histochemical: alkaline phosphatase, lactate dehydrogenase, liver enzymes, creatine kinase, antioxidative enzymes,  - galactosidase. Tests for hemostasis. Cultivation of cells in the suspension. Functional assays for hematopoietic stem cells. Phagocytosis assays. Cell migration assays. The cultivation of adherent cells. Functional tests for mesenchymal stem cells. Determination of cell viability (trypan blue, annexin), methods of cell cycle determination, proliferative activity of cells: MTT and LDH, XTT . Quality control in the laboratory, validation of results, the reference values.
      Contents of exercises All lessons stated under theoretical contents
      Literature
      1. Vunjak-Novaković G. i Freshney RI. Culture of Cells for Tissue Engineering. Wiley-Liss, 2006 (ISBN -13 978-0-471-62935-1); (Original title)
      2. Duncan and Prasse's Veterinary Laboratory Medicine: Clinical Pathology, 5th Edition Kenneth S. Latimer, (ISBN - 978-0-8138-2014-9) (Original title)
      3. Suvarna, K. S., Layton, C., & Bancroft, J.D. (2012). Bancroft’s Theory and Practice of Histological Techniques (7th ed.).New York: Churchill Livingstone (Original title)
      4. Švob M., Histological and histochemical methods. 1974. Svjetlost, Sarajevo
      5. Current Protocols Essential Laboratory Techniques, eds. Sean R. Gallagher and Emily A. Wiley, 2008, (Online ISBN: 9780470089941). (Original title)
      Number of hours per week during the semester/trimester/year
      Lectures Exercises OTC Study and Research Other classes
      4
      Methods of teaching Interactive lectures with the use of multimedia presentations, practical laboratory work, independent experiment.
      Knowledge score (maximum points 100)
      Pre obligations Points Final exam Points
      Activites during lectures 10 Test paper 60
      Practical lessons Oral examination
      Projects 10
      Colloquia 20
      Seminars