Imaging in Pharmacology

Code:

FTEC_14_05

Acronym:

MIF

Keywords
Classification	Keyword
CNAEF	Pharmaceutical Sciences

Instance: 2023/2024 - 1S (of 11-09-2023 to 31-07-2024)

Active?	Yes
Responsible unit:	Departamento de Biomedicina
Course/CS Responsible:	PhD Experimental and clinical Pharmacology and Toxicology

Cycles of Study/Courses

Acronym	No. of Students	Study Plan	Curricular Years	Credits UCN	Credits ECTS	Contact hours	Total Time
PDFTEC	3	Current Studies Plan	1	-	3	14	81

Teaching language

Suitable for English-speaking students

Objectives

To learn the fundamentals of the different imaging techniques in Biology, biological signal processing and the digital analysis of images obtained for studying the relationship between molecular interactions, function and structure of cells and tissues.

Learning outcomes and competences

A picture is said to worth a thousand words but its value increases substantially for pharmacologists with the ability to not only visualize biomolecules but extract quantitative information about their interactions using image analysis, ideally in real time and non-invasively. Imaging sciences have grown exponentially during the past three decades. As a result, image has become an important tool in virtually every branch life sciences. Advances in imaging technologies and imaging probes for humans and for small animals are now extending the applications of imaging further into drug discovery and development, and have the potential to considerably accelerate the process.

Working method

Presencial

Program

High-resolution fluorescence-based imaging (epifluorescence vs confocal microscopy). Instrumentation and detection for live cell imaging; “Hands-on” confocal: Fluorescence protein tracking and detection; monitoring protein dynamics (FRET); binding of fluorescent-labelled ligands; analysis of live cell data; Image capture with increased speed during time-lapse observations while maintaining living cells in the imaging setup; Live cell imaging: high-speed compound application vs photoactivation of caged biomolecules; photobleaching techniques; tracking dynamic cell movements and vesicle exocytosis; Live cell imaging: Single-cell calcium imaging, imaging of voltage-sensitive dyes, real-time synaptic vesicle dynamics using styryl dyes. Laser-capture microdissection of cells from slices and culture dishes.

Mandatory literature

RD Goldman & DL Spector; Live Cell Imaging: A Laboratory Manual, Cold Spring Harbour Laboratory Press, New York
R Yuste & A Konnerth; Imaging in Neuroscience and Development: A Laboratory Manual, Cold Spring Harbour Laboratory Press, New York

Teaching methods and learning activities

Introductory note and practical demonstrations.
Full attendance (±1 week), continuous evaluation and experimental reports.

Developments in imaging receptors both in vitro and in vivo are rapidly progressing with enhancements to both hardware and software which in turn leads to important new methodologies and applications. G-protein-coupled receptors (GPCRs) transduce an eclectic diversity of signals. GPCRs represent targets for nearly half of currently available medicines and continue to be a major focus for the development of novel imaging agents, particularly fluorescent biomolecules. Students will be exposed to techniques of biomolecular fluorescence complementation to GPCRs, yielding measurable binding and functional activity of a given protein (e.g. ion fluxes, second messenger accumulation, homodimerization / heterodimerization). This has the potential to answer key questions as to how GPCRs select downstream signaling partner and how they are compartmentalized within a cell. The great advance in the use of fluorescent labeled ligands is that receptor pharmacology can be visualized and quantified in a single cell. The combined use of confocal imaging with fluorescent labeled ligands increased the special precision of receptor binding. Moreover, fluorescence imaging is not limited to visualize receptors but has the potential to quantify the release and uptake of signaling molecules (e.g. neurotransmitters and mediators). Strategies for non-invasive imaging of gene expression allow its precise spatiotemporal measurement in longitudinal studies. In addition, many pharmacological studies result in the generation of an image of some sort, such as a blot, gel or photomicrograph. With the development of live cell imaging and ultra fast scanning, it is now possible to visualize biological events (e.g. drug responses) in real time. Thus, imaging applied to drug discovery has the unique possibility to bring together high spatial resolution, high sensitivity, and, more recently, high temporal resolution.

Evaluation Type

Distributed evaluation without final exam

Assessment Components

Designation	Weight (%)
Participação presencial	40,00
Teste	30,00
Trabalho laboratorial	30,00
Total:	100,00

Amount of time allocated to each course unit

Designation	Time (hours)
Apresentação/discussão de um trabalho científico	40,00
Estudo autónomo	28,00
Frequência das aulas	14,00
Total:	82,00

Eligibility for exams

Distributed evaluation:

- Practical (6 points): Compulsory attendance (2/3 of attendances). Continuous evaluation by the tutor.

- Theorical and Pratical (8 point): Seminars presentation and discussion.

- Theory (6 points): Final examination (20 multiple choice quiz + 5 short answers) and/or oral examination (students must have >3.0/14.0 to succeed; compulsory between 2.0-2.9/6.0 points in the final examination).

Calculation formula of final grade

[(Written evaluation*30%)+(Theorical/Practical evaluation*40%)+(Pratical evaluation *30%)]/100

Special assessment (TE, DA, ...)

Oral exam.

Classification improvement

Oral exam.