Biotechnological and Chemical Sciences in Diagnostics

Electron Microscopy

Electron Microscopy

Academic year 2024/2025

Sommario del corso

• Course objectives
• Results of learning outcomes
• Program
• Course delivery
• Learning assessment methods
• Support activities
• Teaching tools
• Course card

Course objectives

The teaching contributes to the realization of the educational objectives of the physico-chemical areas of the Master in Biotechnological and Chemical Sciences in Diagnostics, providing ithe basics of the theoretical and practical basis of electronic and atomic force microscopies, important for their applications in the biotechnological field.

The teaching aims to delve into the fundamental properties and stability of colloidal systems, with specific focus on plasmonic nanoparticles and their optical properties, that have interest nanomaterials for molecular diagnostics and therapy applications.

In addition, it provides the students a proper basis for understanding the issues related to the use of electronic and atomic force microscopies to study nanomaterials and biomolecules, as well as a theoretical and practical background to correctly interpret electronic microscopies and AFM images. It also provides basis for a knowledge-driven choice of plasmonic nanoparticles for specific applications.

Results of learning outcomes

At the end of the course the student will be able to:

Knowledge and understanding

- understand the electronic and optical properties of plasmonic nanoparticles, how these are affected by the environment and how these properties can be exploited for technologically important fields such as nanobiotechnology and diagnostics

- know the principles of electron and atomic force microscopies, their potentiality, limitations and fields of application

- know the basics about crystalline materials and the information that can be obtained by diffraction

- know the basics of Circular Dichroism and its usefulness in the study of biomolecules

Apply knowledge and understanding

- read and interpret SEM, TEM and AFM images

- extract quantitative information from SEM, TEM and AFM images (particle size distribution, surface roughness, etc.)

- correctly interpreting the changes in the optical properties of plasmonic NPs as a function of the environment

- analyse CD spectra and extract information on the secondary structure of biomolecules

Making judgements

- Critically choose the proper technique in relation to the desired information

- Be aware of the techniques potentialities and limitations

Communication skills

- Correctly use terms specific to the chemistry of colloids

- Correctly use terms and techincal language specific to electronic and atomic force microscopies

- acquire appropriate skills and tools for processing and presenting experimental data

- know how to work in groups

Learning skills

- To develop autonomous learning skills and self-assessment of their own preparation,
apt to undertake subsequent paths with a high degree of autonomy

Program

The course aims at providing fundamental knowledge about electronic and atomic force microscopies and about the properties of matter at the nanoscale with a specific focus on the stability of nanoparticles in colloidal systems, and how this is affected by their interaction with (bio)molecules. A class of inorganic nanoparticles (plasmonic NPs) with applications in bioimaging and nanomedicine are specifically addressed.

Fundamentals of X-rays diffraction. Use of electronic microscopies for nanoparticles characterization. SEM and TEM. Difference between SE and BSE in SEM images. Interaction volume, resolution, contrast, lens aberrations. Differences between SEM and TEM. Limitations of the techniques. Cryo-TEM. Atomic Force Microscopy. Fluorescence microscopy (hints). 

Analysis of SEM and TEM images.

Inorganic nanoparticles: formation processes (nucleation and growth, aggregation, Ostwald ripening). Nanoparticles stability and flocculation. Metal nanoparticles. Surface plasmons and Localized Surface Plasmon Resonance. Relationship between size/shape/state of aggregation and functional properties of interest for diagnostic applications. Surface Enhanced Resonance Spectroscopy (SERS).

Circular Dicroism.

Laboratory activity about synthesis and characterization of Au nanoparticles. Use of SEM. Data analysis.

Course delivery

The theoretical lectures are in English, supported by presentations (ppt) with graphic illustrations in English. Laboratory activities (6 hours) involve the active participation of the students and include the synthesis and characterization of Au nanoparticles and one session of measurements with SEM.

In addition, one seminar of 2 hours is planned within the teaching, given by an expert in Raman spectroscopy and SERS.

Participation to the lab activities is compulsory.

Learning assessment methods

The exam will be in oral form. The students will be required to interpret SEM, TEM and/or AFM images, explaining the information they can obtain and the basics of the technique (resolution, contrast, lenses aberrations, differences between SEM and TEM, differences between SE and BSE in SEM images, etc.). They will be required to show their understaning about colloidal systems, the properties of plasmonic NPs and/or the basics of Circular Dichroism.

Acquisition and understanding of basic knowledge in Chemistry will be evaluated if necessary in relation to the program, as well as the use of a proper scientific/technical language.

Support activities

Teaching materials (lecture slides, in-depth scientific articles, etc.) related to the teaching is made available before the lectures on Moodle platform. Regarding the practical part of the teaching, laboratory protocols are provided during the experiments and uploaded on Moodle at the end of the activity, so as to include the data obtained in the experiences conducted by the different groups of students/students.

Students are reminded to register for teaching on the Moodle platform so that they can consult the teaching materials and receive any timely communications.
In addition, the lecturer makes herself available to answer any doubts related to the topics of the lectures or laboratory activities, during the lessons or by prior appointment via email (gloria.berlier@unito.it)

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