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Super-resolution fluorescence-assisted diffraction computational tomography reveals the three-dimensional landscape of the cellular organelle interactome

Super-resolution fluorescence-assisted diffraction computational tomography reveals the three-dimensional landscape of the cellular organelle interactome | News Imagerie cellulaire - Cellular imaging | Scoop.it

Dashan Dong, Xiaoshuai Huang, Liuju Li, Heng Mao, Yanquan Mo, Guangyi Zhang, Zhe Zhang, Jiayu Shen, Wei Liu, Zeming Wu, Guanghui Liu, Yanmei Liu, Hong Yang, Qihuang Gong, Kebin Shi & Liangyi Chen

 

Collaborative Innovation Center of Extreme Optics, Shanxi University, Taiyuan, Shanxi, China

and State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing, China


The emergence of super-resolution (SR) fluorescence microscopy has rejuvenated the search for new cellular sub-structures. However, SR fluorescence microscopy achieves high contrast at the expense of a holistic view of the interacting partners and surrounding environment. Thus, we developed SR fluorescence-assisted diffraction computational tomography (SR-FACT), which combines label-free three-dimensional optical diffraction tomography (ODT) with two-dimensional fluorescence Hessian structured illumination microscopy. The ODT module is capable of resolving the mitochondria, lipid droplets, the nuclear membrane, chromosomes, the tubular endoplasmic reticulum, and lysosomes. Using dual-mode correlated live-cell imaging for a prolonged period of time, we observed novel subcellular structures named dark-vacuole bodies, the majority of which originate from densely populated perinuclear regions, and intensively interact with organelles such as the mitochondria and the nuclear membrane before ultimately collapsing into the plasma membrane. This work demonstrates the unique capabilities of SR-FACT, which suggests its wide applicability in cell biology in general.

 

Dong et al. Light: Science & Applications (2020) 9:11

Open Access  : https://doi.org/10.1038/s41377-020-0249-4

PDF : https://www.nature.com/articles/s41377-020-0249-4.pdf

 

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Institut de Biologie Intégrative de la Cellule - Actualités

Institut de Biologie Intégrative de la Cellule - Actualités | News Imagerie cellulaire - Cellular imaging | Scoop.it

Next session of the Imaging Club  :

 

Tuesday 21st of Januaryat

 

11AM

 

Building 21 at the CNRS campus of Gif sur Yvette



  Sylvain Trépout

 

 "Introduction to cryo-STEM tomography: application on the in situ structural analysis of the flagellum attachment zone in T. brucei"

 


Come to discover new imaging technics and to share your experience in imaging, sample preparation and analysis !

 

See you there !

 

 

Claire Boulogne

 CNRS – Institut de Biologie Intégrative de la Cellule

Avenue de la Terrasse – Bâtiment 21

91198 Gif-sur-Yvette Cedex

Tél : +33 (0)1 69 82 46 50

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Dynamic organization of Herpesvirus glycoproteins on the viral envelope revealed by super-resolution microscopy

Dynamic organization of Herpesvirus glycoproteins on the viral envelope revealed by super-resolution microscopy | News Imagerie cellulaire - Cellular imaging | Scoop.it

Publication from Paris-Saclay

 

Frauke Beilstein, Gary H. Cohen, Roselyn J. Eisenberg, Valérie Nicolas, Audrey Esclatine, David Pasdeloup

 

Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris‐Sud, Université Paris‐Saclay, Gif‐sur‐Yvette

IPSIT, Microscopy facility, University of Paris-Sud, Châtenay-Malabry, France

Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia, USA

Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, USA

 

The envelopes of Herpesvirus particles contain a variety of different proteins that allow them to infect specific cell types. An essential core set of these proteins is designed to allow viral entry into the cell after adsorption by binding to specific receptors and ultimately inducing fusion between the viral and cellular membranes in a regulated way through a succession of interactions between receptor-binding and fusion-triggering viral proteins. We have identified here for the first time the localization patterns of these essential proteins at the surface of purified virions and we describe how their localization changes after cell attachment. These results illustrate how the dynamics of viral proteins at the surface of the viral particle could participate in optimizing the all-important process of cell binding and membrane fusion.

 

PLoS Pathog. (2019), 2;15(12) : e1008209.

PubMed PMID: 31790506

OPEN ACCESS  :https://doi.org/10.1371/journal.ppat.1008209

PDF : https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1008209&type=printable

 

 

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In vitro role of Rad54 in Rad51-ssDNA filament-dependent homology search and synaptic complexes formation

In vitro role of Rad54 in Rad51-ssDNA filament-dependent homology search and synaptic complexes formation | News Imagerie cellulaire - Cellular imaging | Scoop.it

Publication from Paris-Saclay

 

Tavares EM, Wright WD, Heyer WD, Le Cam E and Dupaigne P.

 

Genome Maintenance and Molecular Microscopy UMR8126 CNRS, Université Paris-Sud, Université Paris-Saclay, Gustave Roussy, F-94805, Villejuif Cedex

Department of Microbiology and Molecular Genetics, University of California, Davis, Davis, CA, 95616-8665, USA

 

 

Homologous recombination (HR) uses a homologous template to accurately repair DNA double-strand breaks and stalled replication forks to maintain genome stability. During homology search, Rad51 nucleoprotein filaments probe and interact with dsDNA, forming the synaptic complex that is stabilized on a homologous sequence. Strand intertwining leads to the formation of a displacement-loop (D-loop). In yeast, Rad54 is essential for HR in vivo and required for D-loop formation in vitro, but its exact role remains to be fully elucidated. Using electron microscopy to visualize the DNA-protein complexes, here we find that Rad54 is crucial for Rad51-mediated synaptic complex formation and homology search. The Rad54−K341R ATPase-deficient mutant protein promotes formation of synaptic complexes but not D-loops and leads to the accumulation of stable heterologous associations, suggesting that the Rad54 ATPase is involved in preventing non-productive intermediates. We propose that Rad51/Rad54 form a functional unit operating in homology search, synaptic complex and D-loop formation. Homologous recombination uses a template to accurately repair DNA double-strand breaks and stalled replication forks to maintain genome stability. Here authors use electron microscopy to investigate the role of Rad54 in homology search and synaptic complex formation.

 

Pubmed : Nat Commun. 2019 Sep 6;10(1):4058

PMID : 31492866 

Open access :  doi.org/10.1038/s41467-019-12082-z

 PDF : https://www.nature.com/articles/s41467-019-12082-z.pdf

 

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Emmanuel Beaurepaire lauréat du grand prix Raimond Castaing 2019 pour les sciences du vivant

Emmanuel Beaurepaire lauréat du grand prix Raimond Castaing 2019 pour les sciences du vivant | News Imagerie cellulaire - Cellular imaging | Scoop.it

Emmanuel Beaurepaire, chercheur au Laboratoire d’optique et biosciences (Cnrs UMR7645/ Inserm U1182 – Ecole Polytechnique) a reçu jeudi 4 juillet le prestigieux prix décerné lors de la seizième édition du colloque de la Société Française des Microscopies (Sfµ) à Poitiers et qui a rassemblé plus de 320 participants (https://sfmu.fr).
 
L’objet du grand prix Raimond Castaing est de récompenser des chercheurs confirmés ayant contribué de manière remarquable à la microscopie soit en instrumentation et théorie de l’instrument, soit dans l'utilisation innovante d’un instrument en sciences du vivant ou en sciences de la matière.

Emmanuel Beaurepaire a rendu hommage à l’investissement des membres du groupe de microscopie du Laboratoire d’optique et biosciences de l’École polytechnique, à l’origine de nombreuses innovations en imagerie des tissus, en microscopie multiphotonique, en imagerie sans marquage, en imagerie de la biologie du développement et tout dernièrement en imagerie multicouleurs du cerveau avec la microscopie ChroMS.

 

 

Thème de recherche et publications https://portail.polytechnique.edu/lob/en/emmanuel-beaurepaire#CV

 

 Laboratoire

https://portail.polytechnique.edu/lob/fr/recherche/microscopies-avancees-physiologie-des-tissus

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Comparison of Optically‐Derived Biomarkers in Healthy and Brain Tumor Tissue Under One‐ and Two‐Photon Excitation

Comparison of Optically‐Derived Biomarkers in Healthy and Brain Tumor Tissue Under One‐ and Two‐Photon Excitation | News Imagerie cellulaire - Cellular imaging | Scoop.it

 

Sibai M, Mehidine H, Mouawad EK, Jucheaux M, Varlet P, Devaux B, Haidar DA.


Laboratoire Imagerie et Modélisation en neurobiologie et cancérologie (IMNC / CNRS – Universités Paris-Saclay et Paris Diderot).

Centre de Psychiatrie et de Neurosciences, Université Paris Descartes

 

https://onlinelibrary.wiley.com/doi/abs/10.1002/jbio.201900111

The surgical outcome of brain tumor resection and needle biopsy is significantly correlated to the patient's prognosis. Brain tumor surgery is limited to resecting the solid portion of the tumor as current intraoperative imaging modalities are incapable of delineating infiltrative regions. For accurate delineation, in situ tissue interrogation at the submicron scale is warranted. Additionally, multimodal detection is required to remediate the genetically and molecularly heterogeneous nature of brain tumors, notably, that of gliomas, meningioma and brain metastasis. Multimodal detection, such as spectrally‐ and temporally‐resolved fluorescence under one‐ and two‐photon excitation, enables characterizing tissue based on several endogenous optical contrasts. In order to assign the optically‐derived parameters to different tissue types, construction of an optical database obtained from biopsied tissue is warranted. This report showcases the different quantitative and semi‐quantitative optical markers that may comprise the tissue discrimination database. These include: the optical index ratio, the optical redox ratio, the relative collagen density, spectrally‐resolved fluorescence lifetime parameters, two‐photon fluorescence imaging and second harmonic generation imaging.

 

 

J Biophotonics. 2019  5:e201900111.

PubMed PMID: 31276313.

doi : https://doi.org/10.1002/jbio.201900111

 

 

 

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Imagerie-Gif - Atelier de microscopie confocale (lundi 14/10/2019 au vendredi 18/10/2019)

Imagerie-Gif - Atelier de microscopie confocale (lundi 14/10/2019 au vendredi 18/10/2019) | News Imagerie cellulaire - Cellular imaging | Scoop.it

LIEU : Imagerie-Gif Plateforme d'Imagerie Photonique (Gif/Yvette - 91)

OBJECTIFS : 
- Acquérir et renforcer les bases théoriques de la microscopie photonique
- Approfondir et maîtriser l'utilisation pratique d'un microscope plein champ et confocal
- Intégrer les principes et possibilités des nouvelles applications et développements en microscopie photonique
- Comprendre les complémentarités des techniques de la microscopie conventionnelle à la super-résolution


PUBLIC :
Chercheurs, ingénieurs, techniciens


PRÉREQUIS :
Une expérience en microscopie photonique est essentielle.


EQUIPEMENTS/LOGICIELS : Microscopes : Leica SP8 et SP8X, Zeiss LSM 880, Spinning-Disk Roper, Nikon N-STORM, MetaMorph, ImageJ, Huygens déconvolution

 

RESPONSABLES : Romain LE BARS, Laetitia BESSE, Sandrine LECART

 
COÛT PÉDAGOGIQUE : 1600 Euros
 
 
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Light sheet fluorescence microscopy versus confocal microscopy :   in quest of a suitable tool to assess drug and nanomedicine penetration into multicellular tumor spheroids

Light sheet fluorescence microscopy versus confocal microscopy :   in quest of a suitable tool to assess drug and nanomedicine penetration into multicellular tumor spheroids | News Imagerie cellulaire - Cellular imaging | Scoop.it

Publication from Paris-Saclay University

 

Lazzari G, Vinciguerra D, Balasso A, Nicolas V, Goudin N, Garfa-Traore M, Féher A, Dinnyés A, Nicolas J, Couvreur P, Mura S.

 

- Institut Galien Paris-Sud, UMR 8612, CNRS, Université Paris-Saclay, Faculté de Pharmacie, Châtenay-Malabry cedex, France

- Institut Paris-Saclay d'Innovation Thérapeutique (IPSIT), UMS IPSIT Université Paris-Sud US 31 INSERM, UMS 3679 CNRS, Microscopy Facility, Châtenay-Malabry, France

- Structure Fédérative de Recherche (SFR) Necker, Inserm US 24 CNRS UMS 3633, Cell Imaging Platform UMS 24, Hôpital Necker Enfants Malades, Bâtiment Imagine, Paris, France

- BioTalentum Ltd, Gödöllő, Hungary



We recently constructed a multicellular spheroid model of pancreatic tumor based on a triple co-culture of cancer cells, fibroblasts and endothelial cells and characterized by the presence of fibronectin, an important component of the tumor extracellular matrix. By combining cancer cells and stromal components, this model recreates in vitro the three-dimensional (3D) architecture of solid tumors. In this study, we used these hetero-type spheroids as a tool to assess the penetration of doxorubicin (used as a model drug) through the whole tumor mass either in a free form or loaded into polymer nanoparticles (NPs), and we investigated whether microscopy images, acquired by Confocal Laser Scanning Microscopy (CLSM) and Light Sheet Fluorescence Microscopy (LSFM), would be best to provide reliable information on this process. Results clearly demonstrated that CLSM was not suitable to accurately monitor the diffusion of small molecules such as the doxorubicin. Indeed, it only allowed to scan a layer of 100 µm depth and no information on deeper layers could be available because of a progressive loss of the fluorescence signal. On the contrary, a complete 3D tomography of the hetero-type multicellular tumor spheroids (MCTS) was obtained by LSFM and multi-view image fusion which revealed that the fluorescent molecule was able to reach the core of spheroids as large as 1 mm in diameter. However, no doxorubicin-loaded polymer nanoparticles were detected in the spheroids, highlighting the challenge of nanomedicine delivery through biological barriers. Overall, the combination of hetero-type MCTS and LSFM allowed to carry out a highly informative microscopic assessment and represents a suitable approach to precisely follow up the drug penetration in tumors. Accordingly, it could provide useful support in the preclinical investigation and optimization of nanoscale systems for drug delivery to solid tumors.

 

Eur J Pharm Biopharm. 2019?  pii: S0939-6411(19)30301-7

Open acces on doi: https://doi.org/10.1016/j.ejpb.2019.06.019

PubMed PMID: 31228557

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ENDOPLASMIC RETICULUM_2019 — From Basic Cell Biology To Translational Approaches

ENDOPLASMIC RETICULUM_2019 — From Basic Cell Biology To Translational Approaches | News Imagerie cellulaire - Cellular imaging | Scoop.it

Organized with the support of Institute of Psychiatry and Neurosciences of Paris, the Inserm ITMO Cell Biology, Development And Evolution et Avesian

 

Announcement : International conference  « ENDOPLASMIC RETICULUM_2019 — From Basic Cell Biology To Translational Approaches »   will be held from 9 -11th October, 2019 IPNP, Paris.

Registration free at : https://itbcde.aviesan.fr/index.php?pagendx=413


Where : Institute of Psychiatry and Neurosciences of Paris – 102-108, rue de la Santé – Paris 14

Keynote speakers: Göhkan HOTAMISLIGIL, Boston, MA, USA – Gisou VAN DER GOOT, Lausanne, Switzerland

Organizers: Eric CHEVET, Rennes  —  Laurent COMBETTES, Orsay  —  Fabienne FOUFELLE, Paris  —  Thierry GALLI, Paris

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Insight into microtubule nucleation from tubulin-capping proteins

Insight into microtubule nucleation from tubulin-capping proteins | News Imagerie cellulaire - Cellular imaging | Scoop.it

Publication from Paris-Saclay


Valérie Campanacci, Agathe Urvoas, Soraya Cantos-Fernandes, Magali Aumont-Nicaise, Ana-Andreea Arteni, Christophe Velours, Marie Valerio-Lepiniec, Birgit Dreier, Andreas Plückthun, Antoine Pilon, Christian Poüs, Philippe Minard, and Benoît Gigant

 

Institute for Integrative Biology of the Cell , CEA, CNRS,  Univ Paris-Saclay, Gif-sur-Yvette 
Department of Biochemistry, University of Zurich, CH-8057 Zurich, Switzerland
INSERM UMR-S 1193, Université Paris-Sud, Université Paris-Saclay, Châtenay-Malabry
Biochimie, APHP, Hôpitaux Universitaires de l'Est Parisien
INSERM UMR-S 1193, Université Paris-Saclay, Châtenay-Malabry

Biochimie-Hormonologie, APHP, Hôpitaux Universitaires Paris-Sud, Clamart

  

correspondence  : christian.pous@u-psud.fr   --   benoit.gigant@i2bc.paris-saclay.fr.

 


Significance
Microtubules are involved in many key functions of eukaryotic cells, including cell division, intracellular transport, and cell shape. They are hollow tubes made of parallel filaments, themselves formed by the self-assembly of αβ-tubulin molecules. Whereas microtubules lengthen and shorten from their ends dynamically, their birth, called nucleation, remains poorly understood. To gain information on this process, we have determined the structure of tubulin bound to CopN, a bacterial protein that delays nucleation. Together with the behavior of artificial tubulin-binding proteins, our results lead to the hypothesis that targeting two filaments at the fast-growing end of the microtubule inhibits nucleation. They also suggest different dynamics at both ends of the nucleus.

 

 

Proc Natl Acad Sci U S A. 2019 Apr 29. pii: 201813559.

https://doi.org/10.1073/pnas.1813559116

PMID :31036638

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Combining 3D single molecule localization strategies for reproducible bioimaging

Combining 3D single molecule localization strategies for reproducible bioimaging | News Imagerie cellulaire - Cellular imaging | Scoop.it

Publication from Paris-Saclay

 

Clément Cabriel, Nicolas Bourg, Pierre Jouchet, Guillaume Dupuis, Christophe Leterrier, Aurélie Baron, Marie-Ange Badet-Denisot, Boris Vauzeilles, Emmanuel Fort & Sandrine Lévêque-Fort

 

Institut des Sciences Moléculaires d'Orsay, CNRS,Université Paris-Saclay

Centre de Photonique BioMédicale, Université Paris-Saclay, CNRS, Fédération LUMAT

Aix-Marseille Université, CNRS, INP, NeuroCyto, Marseille

Centre de Recherche de Gif, Institut de Chimie des Substances Naturelles du CNRS, Gif-sur-Yvette

Laboratoire de Synthèse de Biomolécules, Institut de Chimie Moléculaire et des Matériaux d'Orsay, Université Paris-Saclay, CNRS.

Institut Langevin, ESPCI Paris, PSL University, CNRS

Institut des Sciences Moléculaires d'Orsay, CNRS, Université Paris-Saclay


Here, we present a 3D localization-based super-resolution technique providing a slowly varying localization precision over a 1 μm range with precisions down to 15 nm. The axial localization is performed through a combination of point spread function (PSF) shaping and supercritical angle fluorescence (SAF), which yields absolute axial information. Using a dual-view scheme, the axial detection is decoupled from the lateral detection and optimized independently to provide a weakly anisotropic 3D resolution over the imaging range. This method can be readily implemented on most homemade PSF shaping setups and provides drift-free, tilt-insensitive and achromatic results. Its insensitivity to these unavoidable experimental biases is especially adapted for multicolor 3D super-resolution microscopy, as we demonstrate by imaging cell cytoskeleton, living bacteria membranes and axon periodic submembrane scaffolds. We further illustrate the interest of the technique for biological multicolor imaging over a several-μm range by direct merging of multiple acquisitions at different depths.

 

Nat Commun. 2019, 30;10(1):1980

https://doi.org/10.1038/s41467-019-09901-8

PMID: 31040275

Open Acces : https://www-nature-com.gate2.inist.fr/articles/s41467-019-09901-8.pdf

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Supercritical Angle Fluorescence for Enhanced Axial Sectioning in STED Microscopy - ScienceDirect

Supercritical Angle Fluorescence for Enhanced Axial Sectioning in STED Microscopy - ScienceDirect | News Imagerie cellulaire - Cellular imaging | Scoop.it

 

Publication from UPSaclay

 

Sivankutty S, Hernández IC, Bourg N, Dupuis G, Lévêque-Fort S.

 

Institut des Sciences Moléculaires d’Orsay, CNRS, Univ. Paris-Sud, Université Paris-Saclay, F-91405 Orsay, France

Centre Laser de l’Université Paris Sud (CLUPS/LUMAT), Univ. Paris-Sud, CNRS, Université Paris-Saclay, F-91405 Orsay, France

 

We demonstrate subwavelength axial sectioning on biological samples with a stimulated emission depletion (STED) microscope combined with supercritical angle fluorescence (SAF) detection. SAF imaging is a powerful technique for imaging the membrane of the cell based on the direct exploitation of the fluorophore emission properties. Indeed, only when fluorophores are close to the interface can their evanescent near-field emission become propagative and be detected beyond the critical angle. Therefore, filtering out the SAF emission from the undercritical angle fluorescence (UAF) emission in the back focal plane of a high-NA objective lens permits nanometer axial sectioning of fluorescent emitters close to the coverslip. When combined with STED microscopy, a straightforward gain in axial resolution can be reached without any alteration of the STED beam path. Indeed, STED-SAF implementation only requires a modification in the detection path of the STED microscope and thus could be widely implemented.

 

Methods 2019  pii: S1046-2023(18)30427-4

https://doi.org/10.1016/j.ymeth.2019.03.027

PMID: 30946895.

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Multicolour single-molecule tracking of mRNA interactions with RNP granules

Multicolour single-molecule tracking of mRNA interactions with RNP granules | News Imagerie cellulaire - Cellular imaging | Scoop.it

Moon SL, Morisaki T, Khong A, Lyon K, Parker R, Stasevich TJ.

 

Ribonucleoprotein (RNP) granules are non-membrane-bound organelles that have critical roles in the stress response1,2, maternal messenger RNA storage3, synaptic plasticity4, tumour progression5,6 and neurodegeneration7,8,9. However, the dynamics of their mRNA components within and near the granule surface remain poorly characterized, particularly in the context and timing of mRNAs exiting translation. Herein, we used multicolour single-molecule tracking to quantify the precise timing and kinetics of single mRNAs as they exit translation and enter RNP granules during stress. We observed single mRNAs interacting with stress granules and P-bodies, with mRNAs moving bidirectionally between them. Although translating mRNAs only interact with RNP granules dynamically, non-translating mRNAs can form stable, and sometimes rigid, associations with RNP granules with stability increasing with both mRNA length and granule size. Live and fixed cell imaging demonstrated that mRNAs can extend beyond the protein surface of a stress granule, which may facilitate interactions between RNP granules. Thus, the recruitment of mRNPs to RNP granules involves dynamic, stable and extended interactions affected by translation status, mRNA length and granule size that collectively regulate RNP granule dynamics.

 

Nat Cell Biol. 2019; 21(2):162-168

Doi : https://doi.org/10.1038/s41556-018-0263-4

 

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Initiation à la microscopie électronique en transmission : de la cellule à la molécule   -  Imagerie-Gif

Initiation à la microscopie électronique en transmission : de la cellule à la molécule   -  Imagerie-Gif | News Imagerie cellulaire - Cellular imaging | Scoop.it

From : Claire Boulogne (UPSaclay)

 

Vous ne le savez pas encore, mais vous pouvez tout faire grâce à la microscopie électronique. Ne ratez pas l’occasion de le découvrir : inscrivez-vous !!!

 

Cette formation organisée avec l’IFSEM aura lieu du 25 au 27 mars à Gif-sur-Yvette, sur les plateformes de microscopie électronique de l’I2BC (cryo-microscopie et Imagerie-Gif).

programme  : ici
inscription  : ici  (deadline 15 février)

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Analyze Microscopy Images - Live Demo

Analyze Microscopy Images - Live Demo | News Imagerie cellulaire - Cellular imaging | Scoop.it

Annonce UPSaclay 

 

Jeudi 13 février de 11h à 12h30 aura lieu une présentation des potentialités du logiciel IMARIS  (lien).

 

La présentation aura lieu à l'hôpital Paul Brousse, au Centre Hépato-Biliaire,  auditorium Paul Barret (plan), situé au 2ème étage

 

Inscription : larbi.amazit@u-psud.fr

 

 

La plateforme d'imagerie de l'UMS 44 (Hôpital Paul Brousse) possède le logiciel IMARIS  qui est dédié à l'analyse d'image.   Il permet notamment, à partir de vos acquisitions, de réaliser des opérations de segmentation, de tracking, de reconstruction volumique et surfacique (3D/4D) et d'édition de vidéos d’animations multidimensionnelles.   Notre station de travail Imaris possède une fonctionnalité maximale avec de nombreux modules dont MeasurementPro, ImarisTrackLineage, ImarisColoc, ImarisXT,ImarisCell,FilamentTracer, ImarisVantage, ImarisBatch, etc...

 

Imaris est en libre-service sur la Plate-Forme d’imagerie de Paul Brousse après réservation sur le site intranet.

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Multimodal imaging for tumour characterization from micro‐ to macroscopic level using a newly developed dorsal chamber designed for long‐term follow‐up

Multimodal imaging for tumour characterization from micro‐ to macroscopic level using a newly developed dorsal chamber designed for long‐term follow‐up | News Imagerie cellulaire - Cellular imaging | Scoop.it

Publication from UPSaclay


Rouffiac V, Roux KS, Salomé-Desnoulez S, Leguerney I, Ginefri JC, Sébrié C, Jourdain L, Lécluse Y, Laplace-Builhé C

 

Gustave Roussy, Plate-forme Imagerie et Cytométrie, UMS 23/3655,, Université Paris-Saclay
Gustave Roussy, Plate-forme d'évaluation préclinique, UMS 23/3655, Université Paris-Saclay
IR4M ,UMR CNRS 8081, Université Paris-Saclay.


Optical imaging of living animals is a unique method of studying the dynamics of physiological and pathological processes at a subcellular level. One‐shot acquisitions at high resolution can be achieved on exteriorized organs before animal euthanasia. For longitudinal follow‐up, intravital imaging can be used and involves imaging windows implanted in cranial, thoracic or dorsal regions. Several imaging window models exist, but none have proven to be applicable for long‐term monitoring and most biological processes take place over several weeks. Moreover, none are compatible with multiple imaging modalities, meaning that different biological parameters cannot be assessed in an individual animal. We developed a new dorsal chamber that was well tolerated by mice (over several months) and allowed individual and collective cell tracking and behaviour analysis by optical imaging, ultrasound and magnetic resonance tomography. This new model broadens potential applications to areas requiring study of long‐term biological processes, as in cancer research.

 

J Biophotonics. 2019 Oct 8:e201900217.

Pubmed PMID: 31593616 

doi :  https://doi.org/10.1002/jbio.201900217

 

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Imaging organoids, spheroids & 3D cell cultures - Sciencesconf.org

Imaging organoids, spheroids & 3D cell cultures - Sciencesconf.org | News Imagerie cellulaire - Cellular imaging | Scoop.it

Imaging organoids, spheroids & 3D cell cultures

 

Institut du Cerveau et de la Moelle épinière

 

Hôpital Pitié Salpétrière

 

Wednesday, 9th October 2019

 

Keynote speakers:

 

Academic partners:

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IMAGERIE   et  SIGNALISATION   --  14 novembre 2019   --  5ème Journée RIC Paris-Saclay  (Faculté de Médecine Paris-Sud)

IMAGERIE   et  SIGNALISATION   --  14 novembre 2019   --  5ème Journée RIC Paris-Saclay  (Faculté de Médecine Paris-Sud) | News Imagerie cellulaire - Cellular imaging | Scoop.it

 

Jeudi 14 novembre prochain aura lieu

la 5ème journée d’imagerie du RIC Paris-Saclay.

 

Trois thèmes : Imagerie ionique - Imagerie de la transcription - Imagerie de la mitochondrie

 

Inscription gratuite mais obligatoire :  ric.inscription@u-psud.fr

 

Lieu : Faculté de médecine Paris-Sud 

 

Programme :  https://bit.ly/33312kr

 

Info : site du RIC

  

 

SAVE THE DATE : 14 NOVEMBRE 2019 -  (9h00 / 15h45)

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AFM-STED correlative nanoscopy reveals a dark side in fluorescence microscopy imaging | Science Advances

AFM-STED correlative nanoscopy reveals a dark side in fluorescence microscopy imaging | Science Advances | News Imagerie cellulaire - Cellular imaging | Scoop.it

Michela Cosentino, Claudio Canale, Paolo Bianchini and Alberto Diaspro

It is known that the presence of fluorophores can influence the dynamics of molecular processes. Despite this, an affordable technique to control the fluorophore distribution within the sample, as well as the rise of unpredictable anomalous processes induced by the fluorophore itself, is missing. We coupled a stimulated emission depletion (STED) microscope with an atomic force microscope to investigate the formation of amyloid aggregates. In particular, we studied the in vitro aggregation of insulin and two alloforms of β amyloid peptides. We followed standard methods to induce the aggregation and to label the molecules at different dye-to-protein ratios. Only a fraction of the fibrillar aggregates was displayed in STED images, indicating that the labeled molecules did not participate indistinctly to the aggregation process. This finding demonstrates that labeled molecules follow only selected pathways of aggregation, among the multiple that are present in the aggregation reaction.

 

Open Acces :

 

Article :  Sci Adv. 2019, 19;5(6):eaav8062

Pumed PMID : 31223651

doi : https://doi.org/10.1126/sciadv.aav8062

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Single molecule super-resolution imaging of bacterial cell pole proteins with high-throughput quantitative analysis pipeline

Single molecule super-resolution imaging of bacterial cell pole proteins with high-throughput quantitative analysis pipeline | News Imagerie cellulaire - Cellular imaging | Scoop.it

publication from Paris-Saclay University


Ipek Altinoglu, Christien J. Merrifield & Yoshiharu Yamaichi

 

Department of Genome Biology and Dpt of Cell Biology, Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, CEA, CNRS, Univ. Paris Sud, Gif sur Yvette, France.

Graduate School of Structure and Dynamics of Living Systems, Univ. Paris-Sud, Orsay, France.



Bacteria show sophisticated control of their cellular organization, and many bacteria deploy different polar landmark proteins to organize the cell pole. Super-resolution microscopy, such as Photo-Activated Localization Microscopy (PALM), provides the nanoscale localization of molecules and is crucial for better understanding of organization and dynamics in single-molecule. However, analytical tools are not fully available yet, in particular for bacterial cell biology. For example, quantitative and statistical analyses of subcellular localization with multiple cells from multiple fields of view are lacking. Furthermore, brightfield images are not sufficient to get accurate contours of small and low contrast bacterial cells, compared to subpixel presentation of target molecules. Here we describe a novel analytic tool for PALM which integrates precisely drawn cell outlines, of either inner membrane or periplasm, labelled by PALM-compatible fluorescent protein fusions, with molecule data for >10,000 molecules from >100 cells by fitting each cell into an oval arc. In the vibrioid bacterium Vibrio cholerae, the polar anchor HubP constitutes a big polar complex which includes multiple proteins involved in chemotaxis and the flagellum. With this pipeline, HubP is shown to be slightly skewed towards the inner curvature side of the cell, while its interaction partners showed rather loose polar localization.

 

Sci Rep. 2019 Apr 30;9(1):6680

https://doi.org/10.1038/s41598-019-43051-7

PubMed PMID: 31040310

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Building bridges to move recombination complexes | PNAS

Building bridges to move recombination complexes | PNAS | News Imagerie cellulaire - Cellular imaging | Scoop.it

Publication from Paris-Saclay University


Emeline Dubois, Arnaud De Muyt, Jessica L. Soyer, Karine Budin, Mathieu Legras, Tristan Piolot, Robert Debuchy, Nancy Kleckner, Denise Zickler, and Eric Espagne

 

Institute for Integrative Biology of the Cell, CNRS, Commissariat à l’Énergie Atomique et aux Énergies Alternatives, Université Paris-Sud, Université Paris-Saclay, 91198, Gif-sur-Yvette

CNRS, UMR3244, Institut Curie, Paris Sciences and Letters Research University, 75005 Paris

UMR Biologie et Gestion des Risques en Agriculture, Institut National de la Recherche Agronomique, AgroParisTech, Université Paris-Saclay, 78850 Thiverval-Grignon

Institut Curie, UMR 3215, INSERM U934, 75005 Paris

Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138


Significance : 

The central feature of meiosis is pairing and recombination of homologous maternal and paternal chromosomes (homologs). Homolog axes become first coaligned at a certain distance; they then synapse by synaptonemal complex (SC) formation. We show that the structural and molecular pathway by which chromosomes transit from coalignment to the SC state in the fungus Sordaria macrospora involves the formation of robust interaxis bridges comprising axis component Spo76/Pds5, recombination proteins, and the evolutionary-conserved Zip2-Zip4 complex. Zip2-Zip4 mediates the recombination complex/structure interface from coalignment onward. These findings solve the conundrum of how recombination complexes move from on-axis localization at coalignment to between-axis localization on SC central regions and provoke new ideas about the molecular and mechanistic nature of SC nucleation.

 

Proc Natl Acad Sci U S A. 2019 May 30 : pii: 201901237

https://doi.org/10.1073/pnas.1901237116

Pumed PMID : 31147459

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Institut de Biologie Intégrative de la Cellule - Imagerie-Gif organizes FLIM-@-GIF (22 May 2PM)

Institut de Biologie Intégrative de la Cellule - Imagerie-Gif organizes FLIM-@-GIF (22 May 2PM) | News Imagerie cellulaire - Cellular imaging | Scoop.it

 

Imagerie-Gif Light Microscopy Facility

 

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Wednesday 22th of May (14h00-17h00) listen to three experts to discover FLIM:


What is Fluorescence Lifetime Imaging Microscopy ?

            Fabienne Mérola (Laboratoire de Chimie Physique, Orsay)

 

Why using Fluorescence Lifetime Imaging in Biology ?

           Sergi Padilla-Parra (Medical Sciences Division, Univ of Oxford)

 

How to perform easy FLIM experiments?

           Clément Laigle (Leica-Microsystems)

 

Inscription is free but mandatory: Inscription link is here!

 

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Thursday 23th and Friday 24th of May: Workshops!


Imagerie-Gif will host for two days the Leica’s SP8 FALCON, an integrated solution for Fluorescence Lifetime Imaging that allows you to:

 

- Follow fast molecular interactions via FLIM-FRET

 

- Use biosensors to detect changes in metabolic state and

   microenvironment

 

- Apply lifetime contrast to separate multiple fluorophores

 

- Acquire fluorescence lifetime data with minimal training

 

Contact us to book a demonstration of the FALCON

and bring your own samples

 

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Contact: imagerie@i2bc.paris-saclay.fr

 

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Practical Course : Expansion Microscopy - 18/19 novembre - Gustave Roussy

Practical Course : Expansion Microscopy - 18/19 novembre - Gustave Roussy | News Imagerie cellulaire - Cellular imaging | Scoop.it

"La microscopie par dilatation"

 

 Cours pratique sur une technique de microscopie récente 

 

"L' Expansion Microsocopy" , c'est quoi ?  : 

 

Applicable sur tout type d’échantillon biologique, cette technique permet l’imagerie de détails très fins en les rendant physiquement plus grands grâce à un processus d'expansion chimique isotrope qui préserve les détails à l'échelle nanométrique (info :  Boyden E.S. 2015 Science)

 

Date : 18 et 19 Novembre

Lieu : Gustave Roussy

Inscription : gratuite mais limitée à 20 personnes

Dead line : 31 mai 2019

 Info et programme : site web de l’évènement.

 Contact : Tudor.MANOLIU - arobase - gustaveroussy.fr

 



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3D + time imaging of normal and twin sea urchin embryos for the reconstruction of their cell lineage 

3D + time imaging of normal and twin sea urchin embryos for the reconstruction of their cell lineage  | News Imagerie cellulaire - Cellular imaging | Scoop.it

 

Publication from UPSaclay

 

Antonio Ortiz, Elena Kardash and Nadine Peyrieras

 

BioEmergences Laboratory (USR3695), CNRS, Université Paris-Saclay, Gif-sur-Yvette, France

 

The Mediterranean sea urchin, Paracentrotus lividus, has been a powerful model to study embryonic development since the late 1800s. As a model, it has the advantage of having external fertilization, it can easily be manipulated experimentally, and it has semi-transparent embryonic stages, which makes it ideal for live imaging. Embryogenesis is a highly dynamic process with intrinsic variability. The reconstruction of cell dynamics and an assessment of such variability from in vivo observations has proven to be a challenge. Here, we provide an innovative methodology for manipulation and immobilization of embryos and their long-term 3D + time imaging. We then describe the twinning procedure that allows us to assess the variability and robustness of developmental processes. We demonstrate the reconstruction of cell lineages based on automated image processing and cell tracking using the BioEmergences workflow as well as the use of interactive visualization tools (Mov-IT software) for lineage validation, correction and analysis.

 

 

Methods Cell Biol. 2019;151:399-418

https://doi.org/10.1016/bs.mcb.2019.01.008

PMID: 30948021.

 

 

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About samples, giving examples: Optimized Single Molecule Localization Microscopy

About samples, giving examples: Optimized Single Molecule Localization Microscopy | News Imagerie cellulaire - Cellular imaging | Scoop.it

Angélique Jimenez, Karoline Friedl, Christophe Leterrier

 

 

Super-resolution microscopy has profoundly transformed how we study the architecture of cells, revealing unknown structures and refining our view of cellular assemblies. Among the various techniques, the resolution of Single Molecule Localization Microscopy can reach the size of macromolecular complexes and offer key insights on their nanoscale arrangement in situ. SMLM is thus a demanding technique and taking advantage of its full potential requires specifically optimized procedures. Here we describe how we perform the successive steps of an SMLM workflow, focusing on single-color Stochastic Optical Reconstruction Microscopy (STORM) as well as multicolor DNA Points Accumulation for imaging in Nanoscale Topography (DNA-PAINT) of fixed samples. We provide detailed procedures for careful sample fixation and immunostaining of typical cellular structures: cytoskeleton, clathrin-coated pits, and organelles. We then offer guidelines for optimal imaging and processing of SMLM data in order to optimize reconstruction quality and avoid the generation of artifacts. We hope that the tips and tricks we discovered over the years and detail here will be useful for researchers looking to make the best possible SMLM images, a pre-requisite for meaningful biological discovery.

 

DOI : http://dx.doi.org/10.1101/568295

Open access :https://www.biorxiv.org/content/biorxiv/early/2019/03/05/568295.full-text.pdf

 

 

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