Dipartimento di Scienze della Vita e dell'Ambiente - Guida degli insegnamenti (Syllabus)
Knowledge of cellular biology, histology, physics and inorganic chemistry
The course includes lectures (4 credits, 32 hours) and laboratory practical activities (2 credits, 16 hours) in which students will learn to make laboratory experiments and to organize the experimental data. Through an online access, students can obtain didactic material and instructions for the laboratory activities.
The course enables students to acquire the fundamental knowledge of the theoretical and methodological basis of main techniques used in a biological laboratory (Centrifugation, spectrophotometric and spectrofluorimetric method, histological and immuno-histochemical analyses, electrophoretic techniques).
Ability to apply the knowledge:
At the end of the course, the student should also acquire the following professional skills: ability to carry out biological laboratory techniques for biochemical, cytological, histological and molecular analyses.
Individual and group laboratory experience improve the student autonomy and the communication capacity for sharing experimental problems. In addition, the use of laboratory equipment provides to the student also transversal skills in biochemistry, cell and molecular biology.
INTRODUCTION TO THE LABORATORY OF BIOLOGY. Codes of conduct and safety precautions in the laboratory, individual protection devices (DPI), use of pipettes and micropipettes equipment usually founded in a laboratory of biology.
PREPARATION OF SOLUTIONS AND BIOLOGICAL BUFFERS. the meaning of the solution concentration, pH definition, preparation and use of the buffer solutions.
SPECTROSCOPIC TECNIQUES. Spectrophotometric and spectrofluorimetric techniques. Components of a Spectrophotometer Spectrum of absorption and analysis of protein concentration by the Lowry method. Enzymatic activities: practical use of spectrophotometric methods to measure the enzyme activities. Uses of fluorophores in biological applications.
OMOGENIZATION, CENTRIFUGATION AND CELL FRACTIONATING. Physical and mechanical methods to homogenate cells and biological tissues. Component of a centrifuge and parameters of a centrifuge run.
LIGHT AND FLUORESCENCE MICROSCOPY TECNIQUES. Light microscope, fluorescence microscope, Transmission electron microscopy (TEM), , Confocal Laser Scanning Microscope (CLSM) Scanning Electron Microscope (SEM). Histological sample preparation for light and electron microscopy. Histochemical and l reactions on micro-tissue sections and staining for the visualization of cellular components
IMMUNOLOGICAL TECHNIQUES. Antibodies production and use in biological application. ELISA-Enzyme Linked Immunosorbent Assay, western blot, immunohistochemical and immunofluorescence techniques.
ELECTROPHORESIS: loading and running the gel for DNA and protein separation; separating protein with SDS-PAGE, 2D-PAGE; separation of DNA molecules with agarose gels.
PRACTICAL ACTIVITY 1. Solutions and biological buffer preparation.
PRACTICAL ACTIVITY 2. Dissection of digestive gland in mussel Mytilus galloprovincialis; homogenization, centrifugation and preparation of cytosolic fractions.
PRACTICAL ACTIVITY 3. Lowry method: colorimetric assay for protein concentration analysis.
PRACTICAL ACTIVITY 4. H&E staining of and lipofuscin staining of cryostat section of digestive gland.
PRACTICAL ACTIVITY 5. DNA gel electrophoresis.
Methods for assessing learning outcomes:
Oral examination in which the students have to answer to the questions (almost 3) concerning the issues of the lectures and the practical activities.
Criteria for assessing learning outcomes:
During the oral exam, the students must demonstrate that they have acquired basic knowledge about the main issues concerning the lessons and practical activities.
Criteria for measuring learning outcomes:
The final mark is assigned in a scale of thirty. The exam is passed when the score is equal or greater than 18. It is expected to be awarded the highest marks with honors (30 cum laude).
Criteria for conferring final mark:
The final score is based on the result of the oral examination of the module of Biology Laboratory A-L and the (written reviewed with public discussion) examination of the module Statistics for Experimental Sciences A-L.
Lecture Notes and slides https://servizi.scienze.univpm.it/moodle
«Biochimica Applicata» M. Stoppini, V. Bellotti EdiSES.