Research Methods in Microscopy

Code:

CB4005

Acronym:

CB4005

Keywords
Classification	Keyword
OFICIAL	Biological Sciences

Instance: 2022/2023 - 1S (of 12-09-2022 to 16-12-2022)

Active?	Yes
Responsible unit:	Microscopy
Course/CS Responsible:	Master in Biochemistry

Cycles of Study/Courses

Acronym	No. of Students	Study Plan	Curricular Years	Credits UCN	Credits ECTS	Contact hours	Total Time
M:BQ	1	Plano de Estudos do MBIOQ_2013-2014	1	-	6	56	162

Teaching language

Suitable for English-speaking students

Objectives

This curricular unit is designed to help students:

- acquire general methodological concepts and knowledge of microscopy techniques.

- be prepared with the essential skills to correctly utilize microscopy techniques in solving specific scientific research challenges.

- be able to execute fundamental laboratory procedures in optical (MO) and electronic transmission (TEM) microscopy, as well as dominate the analysis and interpretation of the obtained data, and be able to replicate them consistently.

Learning outcomes and competences

Following the frequency of this curricular unit, in which students establish direct contact with concrete and updated microscopy methodologies related to laboratory investigation in order to complete their training at the level of knowledge acquired previously either during the tutorial classes or through self-learning (individual study and autonomous outside the contact hours established by the curricular unit), they must:

- acquire skills that can be employed in a variety of Biosciences fields, including scientific research and diagnostics;

- be capable of processing a wide range of biological samples for optical microscopy with varying objectives.

- be able to process diversified biological samples for electron microscopy purposes.

- to master complementary cytological and histological techniques of investigation and diagnosis.

- be able to process and analyze digital images of microscopy.

Working method

Presencial

Pre-requirements (prior knowledge) and co-requirements (common knowledge)

None

Program

The curricular unit is built on a theoretical-practical basis, with theoretical program content coming before practical program content.

1 - The different types of microscopes, their principles, features, and applications:

Bright field, dark background, phase contrast, and differential interference contrast (contrast techniques). Fluorescence, confocal, and two photons (fluorescence techniques). Electronic transmission, scanning, tunnel effect, and atomic force (physical phenomena).

2 - Preparation and processing of biological material for optical and electronic microscopy observation :

Animal experiments and diagnosis require cytological and histological collection, fixation by immersion and perfusion, impregnation and inclusion in paraffin and EPON, microtomy and ultramicrotomy, staining, and double contrast.

3 - Special techniques:

Histochemical approaches in semithin sections for protein, polysaccharide, and lipid localization in OM. Cytochemical techniques to localize nucleic acids, proteins, and polysaccharides in TEM. Methods for detecting enzymatic activity. Negative contrast. Autoradiography. Cryopreservation techniques such as cryofracture. Immuno-cytochemistry. Immunofluorescence. Stereological methods. Cytogenetic techniques.

4- Photografic images:
Training, digital, processing, and analysis.

Mandatory literature

Bozzola John J.; Electron microscopy. ISBN: 978-0-7637-0192-5
Pawley JB; Handbook of Biological Confocal Microscopy, Pawley JB, 1995
Murphy DB and Davidson MW; Fundamentals of Light Microscopy and Eletronic Imaging, 2013
Goldstein J, Newbury D, Joy D, Lyman C, Echlin P, Sawyer L, Michael J; Scanning Electron Microscopy and X-Ray Microanalysis, 2003
Heath O. V. S.; Investigation by experiment
Eaton P and West P; Atomic Force Microscopy, 2010

Comments from the literature

PubMed International Indexed Articles

Teaching methods and learning activities

Blended learning, incorporating both synchronous and asynchronous activities.

Theoretical instruction with material exposition supported by audiovisual aids.

Laboratory instruction to provide students with the opportunity to practice microscopy methods.

Visits to and from outside sources:

- CEMUP: Scanning Electron Microscope

- I3S: atomic force microscope and flow cytometer; confocal microscope with assistance from an Opympus Engineer.

Evaluation Type

Distributed evaluation with final exam

Assessment Components

Designation	Weight (%)
Exame	60,00
Participação presencial	20,00
Trabalho laboratorial	20,00
Total:	100,00

Amount of time allocated to each course unit

Designation	Time (hours)
Estudo autónomo	40,00
Frequência das aulas	56,00
Trabalho laboratorial	28,00
Total:	124,00

Eligibility for exams

In agreement with the regulations of the University of Porto.

Attendance of at least 75% in practical classes.

The legislation's exception conditions are covered.

Calculation formula of final grade

Final exam note X 0.60 + personal attendance X 0.15 + lab work X 0.15

Passing the Microscopy Research Techniques Course requires a final test grade of 9.5 or above.

Examinations or Special Assignments

None.

Internship work/project

There are no plans for an internship or project work.

Special assessment (TE, DA, ...)

Those stipulated in the University of Porto's Pedagogical Evaluation and Guidance Standards, to be held on dates determined by the ICBAS Pedagogical Council.

Classification improvement

The Pedagogical Council will set the dates for the classification improvement of the final exam (appeal exam).

Improvement in classifications in classroom participation and laboratory work are not foreseen by the University of Porto regulations.

Observations

As a result, the evolution of the Covid-19 pandemic, or the UC reading model, adapts to a consensus with the UPorto rules, if necessary.

However, the UC will resume operations with up to eight students registered.