Menù principale
B017004 - BIOTECHNOLOGY IN MEDICINE
Main information
Teaching Language
Course Content
Suggested readings
Learning Objectives
Prerequisites
Teaching Methods
Further information
Type of Assessment
Course program
Academic Year 2016-17
Course year
Second year - First Semester
Belonging Department
Biomedical, Experimental and Clinical Sciences "Mario Serio"
Modulo di sola Frequenza of
Scientific Area
BIO/16 - HUMAN ANATOMY
Credits
6
Teaching Hours
48
Teaching Term
03/10/2016 ⇒ 30/04/2018
Attendance required
No
Type of Evaluation
Giudizio Finale
Course Content
show
Course program
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Teaching Language
ITALIAN
Course Content
The students will acquire the theoretical and experiemental knowledge of the methods of analysis of proteins, mass spectrometry, immunology, microscopy
Suggested readings (Search our library's catalogue)
Molecular biology text books, Immunobiology textbook, international scientific reviews and papers. DIDACTIC MATERIAL GIVEN BY THE TEACHING STAFF.
Learning Objectives
The students will gain the basic knowhow on protein separation and qualitative/quantitative analysis in biological samples: cells, tissues, biological fluids.
Focus on monodimensional and bidimensional electrophoresis techniques associated to DIGE.
Practical lessons will be focused on monodimensional electrophoresis.
At the end of the course students should have knowledge of the principles of mass spectrometry: ionization mechanisms and ion sources, coupling with gas- and liquid- chromatography, main analyzers, as well as tandem mass spectrometry.
The students should be able to use different mass spectrometry instruments, coupled to GC or LC and also to design a qualitative/quantitative mass spectrometric analysis.
The students will gain the basic knowledge about the standard techniques to isolate the various leukocyte subpopulations from peripheral blood. During the course the students will also receive the main biologic information about dendritic and T cells, with particular regard to those aspects related to in vitro culture techniques.
The students will get the basic knowledge about light, transmission electron microscopy, confocal laser scanning and two-photon excitation microscopy. In particular, during the course will be introduce the basic theoretical principles and techniques of preparation of biological specimens for light, transmission electron, laser scanning confocal and two-photon excitation microscopy. Moreover, students will also receive some principles regard cell culture.
Focus on monodimensional and bidimensional electrophoresis techniques associated to DIGE.
Practical lessons will be focused on monodimensional electrophoresis.
At the end of the course students should have knowledge of the principles of mass spectrometry: ionization mechanisms and ion sources, coupling with gas- and liquid- chromatography, main analyzers, as well as tandem mass spectrometry.
The students should be able to use different mass spectrometry instruments, coupled to GC or LC and also to design a qualitative/quantitative mass spectrometric analysis.
The students will gain the basic knowledge about the standard techniques to isolate the various leukocyte subpopulations from peripheral blood. During the course the students will also receive the main biologic information about dendritic and T cells, with particular regard to those aspects related to in vitro culture techniques.
The students will get the basic knowledge about light, transmission electron microscopy, confocal laser scanning and two-photon excitation microscopy. In particular, during the course will be introduce the basic theoretical principles and techniques of preparation of biological specimens for light, transmission electron, laser scanning confocal and two-photon excitation microscopy. Moreover, students will also receive some principles regard cell culture.
Prerequisites
Nothing
Teaching Methods
LECTURES
PRACTICAL ACTIVITY IN THE LAB
PRACTICAL ACTIVITY IN THE LAB
Further information
No information
Type of Assessment
ORAL EXAMINATION
Course program
Protein analysis
The main techniques of protein analysis on fixed/entracted samples will be covered: immunocytochemistry and immunohistochemistry, autoradiography, fluorescence imaging.
Matrix assisted protein array. Immunorealtime PCR. Immunoprecipitation, Yesat 2 hybrid assay, FRET.
Proteomic analysis
Monodimensional and bidimensional electrophoresis techniques, DIGE and Western Blot analyses. Dosage of sample protein content. Examples of research projects based on protein analysis. Antige.
Antibody reaction.
ELISA and Bioplex
Qualitative and quantitative analysis of proteins based on a single and multiplex antibody targets.
Qualitative/quantitative analysis by mass spectrometry.
Ionization techniques, EI, CI, derivatization for GC-MS analysis, database queries, LC-MS, ESI and APCI interfaces.
Analyzers: magnetic sector analyzer, quadrupole, ion trap, time-of-flight, Q-trap, linear ion trap, FT-ICR.
Orbitrap and high resolution analysis, accurate mass measurements, tandem mass spectrometry. Different kinds of mass spectrometry instruments. Nano HPLC and gain in sensitivity, interfaces for nano HPLC.
Laboratory practice on GC-MS, GC-MS/MS, LC-MS, LC-MS/MS, ion traps, MALDI-TOF, MALDI-orbitrap, LC-MS Orbitrap.
1: Isolation of white cells from whole peripheral blood:
This part comprehendS: a) the gradient density stratification of blood cells on ficol and the subsequent harvesting of mononuclear cells, b) the immunomagnetic cell sorting, starting from mononuclear cells, based on the usage of monoclonal antibody, conjugated with paramagnetic beads, specific for the interest surface lineage markers.
2: In vitro differentiation of dendritic cells:
Student will gain the main biologic aspect and the classification criteria of human dendritic cells, and will also gain the in vitro culture methods to obtain myeloid dendritic cells starting from circulating monocytes or from haematopoietic stem cells.
3. T cell culture: The first part of this argument is aimed to recall the basic knowledge about the biologic aspect of T cells activation and proliferation. In the second part of the course the students will gain the practical knowledge and the techniques related to the generation of antigen specific T cell lines and T cell clones.
1. Light and transmission electron microscopy: theoretical principles and preparation of biological samples.
2. New generation light microscopy: confocal laser scanning and two-photon excitation microscopy. Theoretical principles and preparation of biological samples, confocal immunofluorescence techniques. Image analysis.
3. Introduction and basics techniques of cell culture.
The main techniques of protein analysis on fixed/entracted samples will be covered: immunocytochemistry and immunohistochemistry, autoradiography, fluorescence imaging.
Matrix assisted protein array. Immunorealtime PCR. Immunoprecipitation, Yesat 2 hybrid assay, FRET.
Proteomic analysis
Monodimensional and bidimensional electrophoresis techniques, DIGE and Western Blot analyses. Dosage of sample protein content. Examples of research projects based on protein analysis. Antige.
Antibody reaction.
ELISA and Bioplex
Qualitative and quantitative analysis of proteins based on a single and multiplex antibody targets.
Qualitative/quantitative analysis by mass spectrometry.
Ionization techniques, EI, CI, derivatization for GC-MS analysis, database queries, LC-MS, ESI and APCI interfaces.
Analyzers: magnetic sector analyzer, quadrupole, ion trap, time-of-flight, Q-trap, linear ion trap, FT-ICR.
Orbitrap and high resolution analysis, accurate mass measurements, tandem mass spectrometry. Different kinds of mass spectrometry instruments. Nano HPLC and gain in sensitivity, interfaces for nano HPLC.
Laboratory practice on GC-MS, GC-MS/MS, LC-MS, LC-MS/MS, ion traps, MALDI-TOF, MALDI-orbitrap, LC-MS Orbitrap.
1: Isolation of white cells from whole peripheral blood:
This part comprehendS: a) the gradient density stratification of blood cells on ficol and the subsequent harvesting of mononuclear cells, b) the immunomagnetic cell sorting, starting from mononuclear cells, based on the usage of monoclonal antibody, conjugated with paramagnetic beads, specific for the interest surface lineage markers.
2: In vitro differentiation of dendritic cells:
Student will gain the main biologic aspect and the classification criteria of human dendritic cells, and will also gain the in vitro culture methods to obtain myeloid dendritic cells starting from circulating monocytes or from haematopoietic stem cells.
3. T cell culture: The first part of this argument is aimed to recall the basic knowledge about the biologic aspect of T cells activation and proliferation. In the second part of the course the students will gain the practical knowledge and the techniques related to the generation of antigen specific T cell lines and T cell clones.
1. Light and transmission electron microscopy: theoretical principles and preparation of biological samples.
2. New generation light microscopy: confocal laser scanning and two-photon excitation microscopy. Theoretical principles and preparation of biological samples, confocal immunofluorescence techniques. Image analysis.
3. Introduction and basics techniques of cell culture.