Cellular and Molecular Biology Techniques and Applications in Marine Sciences

Code:

CMRM114

Acronym:

CMRM114

Keywords
Classification	Keyword
OFICIAL	Marine Sciences

Instance: 2013/2014 - 1S

Active?	Yes
Responsible unit:	Microscopy
Course/CS Responsible:	Master Degree in Marine Sciences - Marine Resources

Cycles of Study/Courses

Acronym	No. of Students	Study Plan	Curricular Years	Credits UCN	Credits ECTS	Contact hours	Total Time
MCMRM	18	Specialisation in Marine Biology and Ecology 2008	1	-	5	50	135
		Specialisation in Aquaculture and Fisheries 2008	1	-	5	50	135

Teaching language

Portuguese

Objectives

Familiarize the students with various types of techniques used to analyze biological samples in cellular and molecular biology. Remind students about and clarify the theoretical principles that are at the base of each technique, as well as exemplify some of its most common applications in biology. Allow the applicant's learning experience of "know-how" with regard to the techniques used in molecular and cellular biology.

Learning outcomes and competences

At the end of classes, the student must show that he has knowledge, understanding, implementation capacity and competence in the areas relating to various techniques, cytological, histological (microscopy, optics, electronics and fluorescence), cytometry and molecular (as is still qtRT-PCR). One of the objectives is also provide the experience of working in a group for the resolution of problems.

Working method

Presencial

Program

INTRODUCTION.

EXPERIMENTAL MODELS IN SCIENCES OF the SEA AND ANIMAL WELFARE - FISH AS a MODEL.

COLLECTION OF SAMPLES.

PROCESSING FOR MICROSCOPY OPTICS: THE COLLECTION In BLADE: Introduction. Dissection and collection of samples. Criofixacao. Fixation and decalcification. Dehydration. Diaphanization. Impregnation and inclusion. Cutting.

SAMPLE COLLECTION AND PROCESSING FOR MICROSCOPY ELETRÓNICA: Introduction. Dissection and collection of samples. Processing of the tissue. Ultramicrotomia. Coloring and hallmarking. Criotecnicas. Observation under the microscope. Applications.

COLORING TECHNIQUES IN MICROSCOPY OPTICS: Introduction to theory of coloration. Description of some of the most used techniques in histology: Hemateina & Eosin. Argentum Impregnation: method of Bielschowsky's for nerve fibers and axons; Gomori Method to reticular fibers. Masson trichrome for collagen fibrils. Stainings for lipids. Nissl Stainings for the nervous tissue. Weigert Staining for elastic fibers.

HISTOQUÍMICA: Introduction. Preparation of the tissue. Histochemical Methods. Qualitative versus quantitative Analysis. Applications.

IMUNOHISTOQUÍMICA: Introduction. Production of antibodies. Preparation of the tissue. Direct and indirect Method. Visualization of the final products of the reaction (optical microscopy and electronic). Qualitative versus quantitative Analysis. Applications.

IF-POSITIVE CELLS: Introduction. Antigens. Methods for preservation of the antigen. Selection of antibodies. Procedure. Applications. Introduction to confocal microscopy

TECHNIQUES CITOLÓGICAS: Introduction. General Principles of cytology. Handling of samples. Preparation of the samples. Staining of cytological samples. Cytological Observation. Applications.

CULTURE OF CELLS: Introduction to the basic principles. Equipment. Preparation of reagents and equipment. Primary cell Cultures. Subcultures and cell lines. Contamination. Cryopreservation. Quantification. Applications.

STEREOLOGY: Introduction. The importance of quantification. Target Parameters. Measures in organs, cells and organelos.

INTRODUCTION TO MOLECULAR METHODS: DNA. RNA.Transcri dog.Translation. Genetic Code. Protein.

HIBRIDIZAÇÃO IN-SITU: Introduction.Types of probes. Preparation of tissue and pre-treatment of the sample. Revelation of hybrids. Aib and modes of observation. Qualitative versus quantitative Analysis. Applications.

FLOW CYTOMETRY: Pincipios and applications.

THEORETICAL CONCEPTS ABOUT MOLECULAR BIOLOGY: Structure of the gene, exons, introns, promoters, regulatory mechanisms. Molecular Evolution, homology/analogy. Molecular biology Applications examples.

BIOINFORMÁTICA/ LAB RNA: Bioinformatics: introductory Notes; GenBank and Ensembl; Resources; genomic Alignments, conserved Regions; Phylogeny; practical Exercise. Lab RNA: introductory Notes; Extraction of RNA concentration, quality assessment, synthesis of cDNA.

REAL TIME PCR: introductory Notes on the basic principles of the technique. Laboratory Exercise with the RT-PCR. Discussion of the results. Alternatives: advanced methods of molecular expression.

Mandatory literature

Material distribuído pelos docentes.

Complementary Bibliography

Kim S Suvarna, Christopher Layton, John D. Bancroft; Bancroft's Theory and Practice of Histological Techniques, Churchill Livingstone, 2012. ISBN: 0702042269
Cook, DJ; • Cellular Pathology: An Introduction to Techniques and Applications, Scion Publishing Ltd., 2006. ISBN: 9781904842309
Freshney, RI; • Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, Wiley-Blackwell, 2010. ISBN: 9780470528129
Reed, MG Howard, V; Unbiased Stereology (Advanced Methods), Garland Science, 2005. ISBN: 9781859960899
Richard W. Burry; Immunocytochemistry: A Practical Guide for Biomedical Research , Springer, 2010. ISBN: 1441913033
van Pelt-Verkuil, E., van Belkum, A, Hays, JP; • Principles and Technical Aspects of PCR Amplification, Springer, 2008. ISBN: 9789048175796
Keith Wilson and John Walker; Principles and Techniques of Biochemistry and Molecular Biology , Cambridge University Press, 2010. ISBN: 0521516358

Teaching methods and learning activities

Lectures.
Practical Classes.

keywords

Natural sciences > Biological sciences > Biology > Molecular biology
Natural sciences > Biological sciences > Biology > Cell biology

Evaluation Type

Distributed evaluation with final exam

Assessment Components

Designation	Weight (%)
Exame	50,00
Participação presencial	50,00
Total:	100,00

Eligibility for exams

According to the Law and Regulations in force (see Standards of UPorto and ICBAS).

Calculation formula of final grade

Final Note = Note the Final Exam Note of the Project (1) Note the Project (2).

The Final Examination is quoted up to 10 values. Each Project is quoted up to 5 values. The students who do their presentation in English (in project written and oral presentation) will have a bonus of up to 0.3 values per Project.

Examinations or Special Assignments

(1) Each student will have to submit orally 1 scientific Article (5 min), which has helped to solve a problem or a case of diagnosis in marine sciences in the area of cellular or molecular biology. The master will have to engage in that there is no place the presentation of examples that can be considered, in practice, equal.

(2) oral Presentation of 1 a web page dedicated to one of the techniques used during the lessons (cell biology or molecular) in the context of their application in the area of marine sciences (10 minutes). The work is carried out in groups of 4 - on the day of presentation will be raffled who presents. All the elements can be called upon to answer questions relating to the work.

Note: The overcoming of the times of submission will be negatively reflected in the standings.

Classification improvement

Implementation of new final exam, not havedo place improving the component continues (except if the student frquentar of new unit, in the academic year following).

Observations

Invited Speakers: the curricular unit has a tradition to be able to invite annually invited speakers, to make advances in technical themes selected.

Lessons of Eduardo Rocha were materialized, under the tutelage of the course manager, by the Master Fernanda Cristina Rodrigues Malhão Pereira and Licensee Celia Cristina Carreiras Lopes.