Cellular and Molecular Biology Techniques and Applications in Marine Sciences

Code:

RM03

Acronym:

TABCMCM

Keywords
Classification	Keyword
OFICIAL	Marine Sciences

Instance: 2018/2019 - 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	15	Oficial Plan 2018	1	-	5	50	135

Teaching language

Suitable for English-speaking students

Objectives

To get the students to be more aware of 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 to get into a learning experience of "know-how" with regard to selected 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, cell culture and molecular (as primer design and qtRT-PCR).

One of the learning outputs is also to provide the experience of working in a group for the resolution of problems.

Working method

Presencial

Program

INTRODUCTION.

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.

GENO/CYTOTOXICITY TESTS: the comet assay.

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.

WESTERN BLOT: Technique, Theory, Practices, and Trouble Shooting

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.

HEMATOLOGY: Brief introduction to Princíples and Applications.

Mandatory literature

Andreas Hofmann, Samuel Clokie; Wilson and Walker's Principles and Techniques of Biochemistry and Molecular Biology (8th Edition), Cambridge University Press, 2018. ISBN: 131661476X
John Cook, Phil Warren; Cellular Pathology, third edition: An Introduction to Techniques and Applications , Scion Publishing Ltd., 2015. ISBN: 1907904352
Kim S Suvarna, Christopher Layton, John D. Bancrof;; Bancroft's Theory and Practice of Histological Techniques (8th Edition), Elsevier, 2018. ISBN: 0702068640
R. Ian Freshney; Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, 7th Edition, Wiley-Blackwell, 2016. ISBN: 1118873653
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

Complementary Bibliography

Material distribuído pelos docentes.

Teaching methods and learning activities

Lectures | Practical Training.

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	70,00
Participação presencial	5,00
Trabalho escrito	25,00
Total:	100,00

Amount of time allocated to each course unit

Designation	Time (hours)
Elaboração de projeto	2,00
Estudo autónomo	83,00
Frequência das aulas	50,00
Total:	135,00

Eligibility for exams

According to the Law and Regulations in force (see Standards of UPorto and ICBAS). It is compulsory to attend 3/4 of the lectured classes.

Calculation formula of final grade

Final Grade = Grade of Final Exam + Grade Project + Attendance Premium.

The Final Examination is quoted up to 14 points.

Each Project is quoted up to 5 points. The students who do their presentation in English (in the written project and in the oral presentation) will have a bonus of up to 0.5 points.

The Attendance Premium is of 1.0 points x % of attendance (number of presences / number of classes of, generally, 2 hours).

The final grade is from 0 to 20, rounded to the unit.

Examinations or Special Assignments

Project - Oral Presentation 1 a web page devoted to a detailed presentation of the techniques used during classes (cellular or molecular biology), in its application in the field of marine sciences.

A practical example of its application, based on a good reference 1 original article published in indexed international journal should be included.

The work is done in groups of 4 students. The groups CAN NOT present projects based on the same technique, and so there must be an effort of inter-group coordination.

On the day of presentation (a ''tour of the page'') it will be randomly picked whom presents and justifies the protocol and whom justifies the choice of the application example. All groups elements can be called upon to answer questions relating to the work.

Note: Overcoming the presentation time of the project (15 minutes ) will negatively impact on the final grade.

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.

Dr. Fernanda Cristina Rodrigues Malhão and Dr. Célia Cristina Lopes are part of the specialized teaching group.