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

Course specification
Course titleMethods of examination of cells, tissues and organs in physiological and pathophysiological conditions
Acronym14D2I08
AcronymDOCTORAL ACADEMIC STUDIES
Module
Type of studyPhD academic studies
Lecturer (for classes)
  1. dr Milica Kovačević-Filipović
  2. dr Danijela Kirovski
  3. dr Dragan Gvozdić
  4. dr Natalija Fratrić
  5. dr Sunčica Borozan
  6. dr Anita Radovanović
  7. dr Danica Marković
  8. dr Svetlana Milanović
Lecturer/Associate (for practice)
    Lecturer/Associate (for OTC)
      ESPB10.0Status
      ConditionEnrolled semester
      The goalThe 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 outcomeThe 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 lecturesGood 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 exercisesAll 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
      LecturesExercisesOTCStudy and ResearchOther classes
      4
      Methods of teachingInteractive lectures with the use of multimedia presentations, practical laboratory work, independent experiment.
      Knowledge score (maximum points 100)
      Pre obligationsPointsFinal examPoints
      Activites during lectures10Test paper60
      Practical lessonsOral examination
      Projects10
      Colloquia20
      Seminars