The workshops will take place on Monday afternoon, June 30, in the various laboratories in Toulouse, and on Tuesday morning, July 1.
They last 2 hours, and can be repeated in the following time slots: 1:30 – 3:30 pm, then 4 – 6 pm on Monday afternoon, or 9:30 – 11:30 am on Tuesday morning.
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Coordinators: Claudie JOSSE – Teresa HUNGRIA, Centre Raimond Castaing
Description: Focused ion beams (FIB) coupled with scanning electron microscopy (SEM) have become essential tools for 3D imaging in fields as varied as materials science, semiconductors and biology. Their ability to reveal the fine structure of samples with great precision makes them the technologies of choice for exploring complex materials at nanometric scales.
The principle is to make successive abrasions of a zone of interest using an ion beam, often gallium (but xenon or oxygen can also be used), and to image each cross-section using scanning electron microscopy.
A stack of images is thus produced. Specific image processing software then reconstructs the volume of material.
The workshop will be divided into two parts. The first will be theoretical, with an oral presentation detailing the principles of 3D FIB-SEM tomography, including its operation, applications and advantages. The second part will be experimental, directly in front of the equipment. Participants will be able to observe the cutting process in real time, then manipulate the equipment themselves to put the concepts into practice.
Equipment used: FEI HELIOS nanoLab 600i SEM/FIB
Location and times: Monday June 30, 1:30 – 3:30 pm and Tuesday July 1, 9:30 – 11:30 am, Castaing platform
Number of participants: 5 participants maximum per session
Coordinator: Florent HOUDELLIER, CEMES
Description: Presentation of the “charged particle optics chair” project. Between INSA, Orsayphysics and CNRS, we have set up a new training program dedicated to the development of new instruments using charged particles, such as the FIB, SEM or TEM. This training is part of the INSA engineers’ cycle in the physics department. I propose here an introductory workshop to this activity. We will address the following questions:
- What is charged particle optics and why teach it?
- What calculation tools do we use? We’ll be demonstrating SIMION and EOD (Electron Optics Design) commercial software.
- What practical tools do we use to test our new optics?
- What optics are used in FIB, SEM and TEM and what are their specific features (electrostatic vs. magnetostatic, rotational symmetry vs. multipolar, straight vs. curved optical axis, etc.)?
Equipment used: Presentation of practical work and demos on SIMION and EOD software. Instrumentation room and OPC room. Disassembly of electrostatic and magnetostatic optical elements (lenses, spectrometers, multipoles).
Location and times: INSA physics department, instrumentation room and OPC room, Monday June 30, 4 – 6 pm, and Tuesday July 1, 9:30 – 11:30 am
Number of participants: 10 participants maximum per session
Coordinator: Frédéric MOMPIOU, CEMES
Description: This workshop is aimed at people interested in discovering in-situ tensile testing in a TEM to study deformation mechanisms in materials. We will begin by demonstrating the tools needed to carry out a deformation micro-test in the microscope, in terms of object holders (traction at different temperatures, nano-indention) and sample preparation. We will discuss the merits of the various approaches. We then propose to carry out a tensile experiment on an electro-polished metal alloy. We will explain how conventional TEM observations can provide quantitative information on deformation mechanisms at the sub-micron scale. To do this, we’ll be using the free pycotem software, which enables us to work end-to-end on the analysis of microstructural defects (dislocations & interfaces) from images and diffraction patterns.
Equipment used: MET JEOL2100HC, tensile specimen holder, pycotem software
Location and times: CEMES, Monday June 30, 1:30 pm – 3:30 pm and Tuesday July 1, 9:30 am – 11:30 am
Number of participants: 6 participants maximum per session
Coordinator: Robin COURS, CEMES
Description: FIB/SEM double-beam microscopes are now widely used to produce samples that can be observed by transmission electron microscopy. The beam of focused ions, generally gallium ions, has a resolution of around ten nanometers, and enables the preparation of thin slides with a thickness of less than 50nm on certain materials. Today’s dualbeam systems easily achieve electron transparency of the sample. However, to obtain an extremely thin sample while maintaining high crystalline quality, the operator must optimize the use of electron and ion beams by skilfully varying certain parameters (high voltage, current, scanning speed, angle of attack…).
This workshop will begin with a live demonstration of the various stages involved in preparing an ultrathin slide, ideal for techniques such as HRTEM or EELS observation. The aim will be to show how to combine speed of execution (less than 2 hours on a silicon substrate) with the high precision required to obtain this type of sample.
It will also be an opportunity to discuss FIB preparation and the problems it can pose, depending on the material.
Equipment used: FIB/SEM ThermoFisher Helios NanoLab600i
Location and times: CEMES, Monday June 30, 1:30 pm – 3:30 pm and Tuesday July 1, 9:30 am – 11:30 am
Number of participants: 4 participants maximum per session
Coordinators: Stéphanie BALOR, Célia PLISSON-CHASTANG, Ramteen SHAYAN, Vanessa SOLDAN, CBI
Description: This workshop will introduce participants to the cryo-MET technique. During the workshop, participants will learn how to prepare samples by cryo-plunging, how to mount grids on suitable supports, and how to use high-throughput/high-resolution TEM imaging at low electron doses to acquire cryo-TEM images. After demonstrating how to collect images, participants will be shown the main principles of 3D reconstruction of objects of interest using SPA isolated particle analysis and/or electron tomography. The aim of the workshop is to provide participants with an overview of cryo-TEM and the analysis of the images obtained, in order to understand how this technique has revolutionized structural biology, at both molecular and cellular levels. These cryopreparation techniques are also applicable to the fields of chemistry and materials science. (polymers, nanoparticles, etc.)
Equipment used :
- Automated grid vitrification Leica EMGP
- Talos Arctica high-throughput/high-resolution TFS cryo-microscope, energy loss filter and Gatan BioQuantum and K3 direct electron detection camera.
- GPU computing station for 3D reconstruction from cryo-EM images (LinuxVixion).
Location and times: Centre de Biologie Intégrative – Plateforme METi, Monday June 30, 1:30 – 3:30 pm, then 4 – 6 pm
Number of participants: maximum 4 participants per session
Coordinators: Renaud POINCLOUX, Myriam RAZOUK, Merzouk ZIDANE, IPBS
Description: We will present a new method, called compression force microscopy, which involves analyzing the forces applied by macrophages phagocytosing polyacrylamide micropillars of controlled size and rigidity. We will explain how the pillars were designed, show how deformations are measured and analyzed, using an automated procedure and finite element-based mechanical simulations.
Equipment used: Spinning Disc confocal microscope
Location and times: CBI or IPBS, Monday June 30, 1:30 pm – 3:30 pm and Tuesday July 1, 9:30 am – 11:30 am
Number of participants: 5 participants maximum per session
Coordinators: Armel DESCAMPS-MANDINE, Arnaud PROIETTI, Centre Raimond Castaing
Description: This workshop introduces two complementary techniques for characterizing crystalline orientations. One is a SEM technique derived from EBSD analysis (t-EBSD) and the other is implemented in a transmission electron microscope (STEMx). For this purpose, the same type of thin slide will be studied in two different workshops: on a FEG SEM and on a FEG TEM.
t-EBSD (transmission EBSD) analysis is an EBSD (Electron BackScatter Diffraction) analysis performed in a SEM, on a thinned slide for transmission electron microscopy. The advantage of working in transmission is that crystallographic information can be obtained on grains as small as 15 nm (whereas the lateral resolution is around 100 nm in conventional EBSD, i.e. on a solid sample). The results provide information on crystallographic orientation, grain shape and size, and even textural information. The aim of this part of the workshop is to familiarize participants with the technique, data processing and discuss the complementarity with transmission electron microscopy analysis.
The STEMx is the 4DSTEM diffraction system from Ametek-Gatan. It collects diffraction data on the camera synchronized with conventional STEM imaging (here from JEOL). Once all the diffraction patterns have been collected for each point of the STEM image, virtual masks can be manipulated to highlight certain areas. In this way, virtual BF/DF images can be obtained. This part of the workshop includes a description of the procedure coupled with live observation, and a discussion of data manipulation (use of virtual masks, python code, etc.).
Equipment used: MEB FEG JEOL 7100F equipped with an EBSD camera and on a MET FEG 2100F equipped with the STEMx system (Gatan).
Location and times: Centre Raimond Castaing, Monday June 30, 1:30 – 3:30 pm, then 4 – 6 pm
Number of participants: 10 participants maximum per session
Coordinators Etienne DAGUE, LAAS-CNRS, Childérick SEVERAC, RESTORE
Description: Atomic force microscopy (AFM), developed in 1986, has now become an essential technique for studying living biological samples on a nanometric scale, observed directly in a liquid medium. The operating principle of the AFM is based on a lever, at the end of which is a very fine tip, which comes into direct contact with the sample to either produce high-resolution images, or to probe the mechanical and adhesive properties of the samples.
The first part of this AFM discovery workshop will involve high-resolution imaging of mammalian cells immobilized on a support. To achieve this, a coupling between optical microscopy and AFM will be set up, enabling areas of interest to be targeted optically and then scanned with AFM.
In the second part, we will carry out force measurements to determine the nanomechanical properties of the cells, i.e. their stiffness, or resistance to compression by the tip. The basic principles of nanomechanics will then be explained, enabling participants to discover the full potential of AFM for this type of measurement.
Finally, a discussion of other AFM modalities (specific interaction measurements, functionalization of AFM tips, coupling to microfluidics) will complete the workshop. This workshop is aimed at anyone interested in discovering this technique applied to life sciences, and no basic knowledge is required.
Equipment used: AFM Nanowizard 4 XP, Bruker
Location and times: LAAS-CNRS, Monday June 30, 1.30-3.30 pm and Tuesday July 1, 9.30-11.30 am
Number of participants: 6 participants maximum per session
You can take part in up to 3 workshops during the congress.
You must first pay a workshop fee, then select the workshops you wish to attend.