Principles and applications of fluorescence microscopy
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Author: Alban Belloir
The use of fluorescence microscopy (wide field, confocal, multiphoton, and now superresolution) in combination with genetically encoded fluorescence probes comprise a powerful set of scientific tools to study live cells. However, surprising little practical and theoretical training in such methods exists within standard curricula, particular at the early stage of training (Masters or Doctorate level). This course offers to cover basic optics principles necessary to understand the origin of microscope resolution and design. Participants will get hands-on experience implementing simple optical configurations to illustrate these fundamental principles. Subsequently, participants will perform experiments on state-of-the-art imaging equipment provided by microscope vendors.
Lectures will cover, in depth, the principle behind traditional high resolution imaging methods such as confocal, multi-photon, and the recently developed super-resolutions methods. Students will also learn about fundamental properties of synthetic and genetically encoded fluorescence indicators used for cellular morphology imaging and signaling recording. Finally guest lectures will demonstrate applications of some of the discussed methodologies.
This course is intended for Master’s students and international PhD students who expect to be using advanced live cell fluorescence microscopy for future studies.
Master 2 students from partner Universities (Sorbonne University and University of Paris Cité) MUST NOT register online and must submit their registration request DIRECTLY to contacts in charge of their university course.These students have free access to the course.
All other students must register online and pay their registration fees.
As the 2023 edition of the France-BioImaging Image Contest admissions is still running, we wanted to highlight our previous winners and their projects. Here is a quick throwback to our 2022 winners.
Before getting to the heart of the matter, we want to remind you that you still have time (before November 10th) to submit your best images and try to win your registration fees for one 2024 microscopy-related event! Please make sure you upload your images on the following link:
Last year, we enjoyed the winning images submitted for their artistic take and their quality. Thanks to Carole SIRET, Magalie BENARD and Frédéric FERCOQ for their beautiful images!
1st Place: Carole SIRET, Van de Pavert Team, Centre d'Immunologie de Marseille-Luminy
"Little Monster"
The embryonic formation of lymph nodes, small organs essential for the immune response, is now known. Using light sheet microscopy, scientists were able to determine the dynamics at work in this 13.5-day-old mouse embryo. In blue, the lymphoid cells (LTi), derived from the haematogenous endothelium, a specific tissue of the embryo. They pass into the liver where they proliferate before migrating through the body to give rise to lymph nodes. The 3D information obtained thus makes it possible to follow the interactions of lymph nodes with their environment, in particular with nerve cells, in green, and blood vessels, in white. The lymphatic endothelial cells and some macrophages are visible in red.
Lightsheet Microscopy
Carole Siret is a Research engineer, expert in Lightsheet microscopy, at the Centre d’Immunology Marseille Luminy (CIML) since 2018. She is working in Dr Serge van de Pavert team where they study immune system development. They are particularly interested in the lymph nodes (LN) formation during mouse embryogenesis.
The image she submitted is a projection from a lightsheet acquisition on the UMII (Miltenyi). This image illustrates an E13.5 mouse embryo stained for neurons, LTi (Tissue inducer cells which are the precursor cells for the lymph node), lymphatic and blood vessels. This acquisition was done in the context of the study of the role of Cxcl12 in embryonic LN formation. From previous work it is clear that Cxcl13 and Ccl21 are not expressed present near blood vessels, but it likely that some chemokines, possibly Cxcl12, could be expressed on the endothelial cells. We focus on Cxcl12 since this chemokine has shown to be important for the attraction of several hematopoietic cells. Although it was shown that the receptor for Cxcl12, Cxcr4, is expressed by the mature hematopoietic inducer cells, it is not clear whether it also expressed by the progenitor hematopoietic inducer cells. Next to the possible attraction of hematopoietic cells towards the lymph node anlagen, Cxcl12 is involved in the attraction of nerve fibers. Therefore, the possible role of Cxcl12 could be to both attract hematopoietic cells as well as nerve fibers to initiate a region which is permissive to form lymph nodes.
Thanks to the France-Bioimaging Image Contest, Carole participated to the SFI Congress, where, this year, it was a special joint conference both between the Société Française d’Immunologie (SFI) and the Deutsche Gesellschaft für Immunologie (DGfI). It was a great opportunity to exchange with people at the cutting edge of the immunology field.
2nd Place:Magalie BENARD, Plateforme de Recherche en IMAgerie CEllulaire de Normandie (PRIMACEN), Research infrastructure HeRacLeS, Inserm US 51, CNRS UAR 2026,
"The communication link with others"
Image of a cellular interconnection between two human tumor cells whose cytoskeleton has been labeled with anti-tubulin (ATTO-647N), anti-vimentin (AlexaFluor594) antibodies and with Phalloidin probe (AlexaFluor488). Scale bar 1µm.
Confocal microscopy
Magalie Bénard is a Research Engineer and the Technical Manager at the Cellular Imaging Facilty PRIMACEN (Plate-forme de Recherche en IMAgerie CEllulaire de Normandie).
The image she submitted is a confocal image representing a cellular interconnection tunneling nanotube (TNTs) between two human tumour cells. In a cancer case, some cells are able to express spontaneously TNTs with cytoskeleton protein composition corresponding to specific role of this communication mechanism. In the winning image, the TNT is composed of tubulin (magenta), actin (cyan) and vimentin (yellow) proteins. Called TNT1, this nanotube allows the transfer of intracellular elements such as RNA, proteins or organelles. Moreover, due to the thinness of TNTs, their photo-sensitivity and their fragility, live-cell imaging is technically challenging with regards to potentially damaging methods. Magalie and her team have developed an adapted method to observe TNTs in living cell with high resolution imaging (STED) enhanced by FLIM by using red and near infrared probes.
France-Bioimaging sponsored her participation to the ELMI (European Light Microscopy Initiative Meeting June 6-9, 2023) congress. During this event, she had the chance to present her project through a poster. This congress also offered a great opportunity to have an overview and the last updates on state-of-the-art imaging techniques.
3rd Place:Frédéric FERCOQ, Parasites et Protistes Libres (PPL), Museum National d'Histoire Naturelle
"Sepia"
Stage 25 cuttlefish embryo (Sepia officinalis) observed under a confocal microscope. The cuttlefish was cleared and the tissue autofluorescence was captured.
This image was produced in collaboration with Laure BONNAUD-PONTICELLI and Luis MOLINA from the BOREA laboratory.
Confocal microscopy
Frédéric Fercoq is a postdoc scientist in the Parasitology laboratory of the Muséum National d’Histoire Naturelle (MNHN) in Paris. My main interest is on how myeloid cells participate to the control of parasitic infections, but sometimes at the price of collateral tissue damage. This project involves a lot of microscopy of immune cells, parasites and host tissues to analyse the complex interactions taking place at the site of infection.
The image he submitted has nothing to do with his main project! As he has the chance to work on very different topics and models, this image was acquired as a proof of concept for imaging full embryos of the cuttlefish Sepia officinalis for Frédéric's colleagues Laure BONNAUD-PONTICELLI and Luis MOLINA (BOREA laboratory, MNHN). They work on the nervous system of cephalopod and on the influence of environmental factors during its development. They are now optimizing fluorescent staining for neuronal markers to test the effect of light on the nervous system in situ.
France-Bioimaging sponsored his participation to the FOM (Focus on Microscopy) 2023 congress in Porto. He had the chance to be granted the opportunity to both present his current project through a poster and to give an oral presentation. He was also amazed by the new avenues opened up by the cutting-edge imaging techniques presented throughout the conference.
You can now apply for a new call to get free open access to instruments and services at one of our nodes’ core facility!
We are happy to announce the first Open Call from the Horizon Europe-funded canSERV project! Cancer Researchers are invited to apply for FREE state-of-the-art services and training at several European Research Infrastructures, including Euro-BioImaging ERIC. Within this project, 28 Euro-Bioimaging Nodes (- Yes, France-BioImaging is part of it too! -) offer access to their expertise. It’s an amazing opportunity for the cancer research community to access a wide-ranging portfolio of services.
CanSERV invites applications covering the entire range of the oncology developmental pipeline, from supporting discoveries in fundamental research to translational science and personalized oncology, addressing at least one of the four strategic goals of the Cancer Mission:
Understanding of Cancer
Prevention and Early Detection
Diagnosis and treatment
Quality of life for patients and their families
Researchers from academia, industry and SMEs may apply for several service categories including:
disease models,
cutting-edge imaging and structural biology technologies,
biomarker research and development,
new therapeutic solutions,
complex clinical trial design and support,
personalized oncology implementation pipelines and recommendations and
regulatory support and tools to analyse the socioeconomic dimension of research activities.
Launch of the first call is today, Thursday, October 12, 2023.
Submission deadline is January 4, 2024 at 14.00 CEST.
To know which Euro-Imaging’s technologies can help your research project, visit their website www.eurobioimaging.eu/service
In order to answer image data analysis demands, France-BioImaging is launching its first data machine learning competition! The idea? To develop powerful methods that will then end up in creating standards & benchmarks in the field of bioimaging.
The FBI challenge is hinged on a double contribution: from core facilities engineers and from data scientists. The first group will acquire a large number of images to build a dataset, that will later be used by the algorithms. These images will be produced by microscopy engineers & technicians from FBI’s platforms. As for the second contribution, this is where the challenge starts! The challenge will be later published to have a maximum of data scientists to work on the algorithms that best fulfill the analysis task.
The first project is also based on four pillars:
Open source + FAIR (Findable, Accessible, Interoperable, Reusable)
Supervised learning, it involves annotated datasets to maintain control over performances.
In silico annotations, a computer labeling method to avoid manual annotation and its drawbacks.
Image-to-image analysis tasks, an image analysis tasks which aim to predict an output image from the input one.
We need you!
The first challenge is still under preparation. It will predict the fluorescence image of cell culture from a bright field, phase contrast and/or DIC microscopy label-free image. Nevertheless, we need you to contribute to the dataset that will supply the machine learning! The more images we have, the better the prediction algorithm will be.
The new edition of the Maxime Dahan tribute symposium will take place on December 14-15, 2023 at the Research Center of the Institut Curie, in Paris. The theme of the symposium will be “Manipulation of cellular organization by optogenetics and magnetogenetics”.
With an ever-growing set of tools and techniques to visualize cellular and sub-cellular organization, microscopy is broadly used to decipher how cells function. However, in addition to passive observation, two techniques —optogenetics and magnetogenetics— offer unique possibilities to directly perturb and control this organization using light and magnetism, yielding opportunities to study cell biology from a radically different perspective.
Maxime Dahan was a pioneer in the development and application of optogenetics and magnetogenetics. This 3rd symposium in his memory will bring together scientists from diverse disciplines to share their work and continue his legacy in this exciting field.
The symposium will take place in the Burg amphitheater, at Institut Curie, Paris. Please find attached the flyer and the program.
The symposium will be the occasion to deliver the Maxime Dahan Prize for Innovation in Methods and Instrumentation at the Interface of Physics, Biology & Medicine.
There will be 3 sessions of selected short talks. If you want to submit an abstract, please send it to Aida.Fakhr@curie.fr
Deadline for registration and abstract submission: November 17
Biogenouest has the pleasure to invite you for Gen2Bio 2024 at the Palais du Grand Large in Saint-Malo!
Gen2Bio, the annual biotech congress organized by Biogenouest, is aimed at all biotechnology stakeholders: researchers, engineers and doctoral students in life and environmental sciences, innovative biotech companies, laboratories or research centers, competitiveness clusters, technology parks and technical centers, players in valorization, etc.
In particular, you can discover the service offerings of the network’s 38 technological platforms and can thus follow innovations in life sciences in the West side of France.
In 2024, the 2-day formula will offer 5 plenary conferences, 24 technological workshops divided into 6 sessions, an exhibition space on a stand for our partners and a BtoB space.
FBI opens a call for the recruitment of its next Deputy Director for Internal Affairs
Mission
The Deputy Director for Internal Affairs will help infrastructure management team to oversee several of its key missions: implement a strategic vision for France-BioImaging, encompassing our three pillars, Innovation, Training, Access; develop a roadmap for next generation imaging technologies and services; increase funding opportunities and develop a sustainable cost model; update and improve infrastructure communication goals and actions; develop ours links with the private sector. The Deputy Director will work closely with the Manager of Internal Affairs to tailor specific activities, prepare strategic decision-making and prioritize needs for initiatives.
Mandate of the Deputy Director for Internal Affairs
Help to develop and implement the strategic vision of FBI’s core activities, in Innovation, Training and Services; synthesize and prepare elements to assist in decision-making
Advise the management team to improve cooperation across FBI geographical Nodes; promote the scientific and structuring activities of the infrastructure at the national level, to seek out and federate potential partners, initiate and manage national cooperation programs, and provide expertise and advice.
Develop a roadmap for technological developments and their implementation as FBI services
Help to develop funding opportunities and develop a sustainable cost model for Innovation and Services
Develop links with the private sector, as users of FBI services or partners in FBI core activities including methodological innovations
Organize a prospective reflection on the development of new initiatives to be carried out to improve FBI communication
Skills
Senior scientist (DR or equivalent) with a strong motivation and willingness to strengthen FBI core activities in Innovation, Service and Training;
Knowledge of the national landscape in biological imaging, including in innovation and services;
Experience in the management of research units or facilities, and projects in the science and innovation domain;
Demonstrated experience in fund raising operations for research and innovation projects in biological imaging;
Knowledge of the national and international research infrastructures in Biology and Health environment
Experience in interdisciplinary scientific and stakeholder networking;
Demonstrated ability to work collaboratively with diverse stakeholders;
Excellent communications and public relations skills;
High proficiency in English language
Organisation
France-BioImaging is a national research infrastructure distributed throughout France that provides researchers with access to the latest innovations in biological imaging and aims to accelerate the transfer of technological and methodological innovations in biological imaging to the 22 platforms that constitute the infrastructure. The Deputy Director for Internal Affairs will contribute to the development and strengthening of France-Bioimaging. The Deputy Director for Internal Affairs will work within the national coordination of the France BioImaging infrastructure. He/she will work within a team distributed between Montpellier and Bordeaux, composed of 4 people: the Scientific Director and the Deputy Director for International Affairs, the Internal Affairs Manager, and the External Affairs Manager.
Time Commitment
FBI’s Deputy Director for Internal Affairs should commit approximately 20% of his overall working time to this mission. Virtual coordination meetings (Directors + Managers) occur weekly, and the FBI Executive Board meets virtually monthly. FBI steering committee meets bi-annually. The Deputy Director is appointed for a 5-year mandate and is eligible to renew his service.
Requirements
Frequent travels in France and abroad.
Entry into function is planned as soon as possible / November 15th, 2023.
You can now apply for two new calls to get free open access to instruments and services at one of our nodes’ core facility!
AgroServ is a transdisciplinary project funded by the European Union under the Horizon Europe programme, running until 2027. It supports the research community by funding interdisciplinary agroecology research projects. The supported research will contribute towards sustainable and resilient agri-food systems, exploring topics like plant biology, water, soil, and microorganisms. Euro-BioImaging is a partner in the AgroServ consortium along with 12 other research infrastructures and partners in 23 countries. Nine Euro-BioImaging Nodes offer access – including France-BioImaging – to their expertise and services within this project.
AgroServ has launched the pre-proposal phase of the first Open Call for projects. Selected projects will receive funding for access to integrated research infrastructure services & expertise.
We encourage all researchers who would like to harness the power of imaging and image data to boost their agroecology research project to look at AgroServ’s impressive Catalogue of Services.
Only core facilities from our Ile-de-France Sud node are available through this call. The node will be happy to welcome you for your research project! For more information about the microscopy techniques you can use thanks to this call, please send an email to contact@france-bioimaging.org
Within the framework of the COMULISglobe project, Euro-BioImaging will be awarding a total of 4 grants for access to Euro-BioImaging Nodes – including France-BioImaging! -, specifically designed to support correlative and multimodal imaging.
Two grants will be awarded to European researchers (including from non-EU countries that are Euro-BioImaging members) accessing services at Euro-BioImaging Nodes. Each grant will provide up to EUR 2000 in support of imaging projects that tackle important research questions using two or more imaging modalities.
Two grants will be awarded for international (non-European) researchers accessing services at Euro-BioImaging Nodes. Each grant will provide up to EUR 3000 in support of imaging projects employing two or more imaging modalities.
This event, organized by the Bioimaging axis and the Image analysis working groupof Biogenouest, will take place at Pôle Numérique Rennes Villejean, on October 4th, 2023.
The Advanced Light Microscopy initiative is organizing a technical symposium involving several microscopy techniques either emerging or identified as lacking awareness from a wider audience. This symposium will take place at the Institut Pasteur in Paris on the 26th of October 2023. This day will orbit around stimulating exchanges between system building scientists and Life Science Researchers in need of new insight to their current available possibilities.
The confirmed speakers as well as the final program will be available soon.
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