Co-localisation studies using three-colour 3D dSTORM imaging with Everspark 2.0

Centrosomal proteins Cep152, rootletin and PCM labelled with AF647, CF568 & AF488 respectively were imaged on a Vutara VXL (Bruker) in Everspark 2.0 buffer before 3D dSTORM reconstruction
Credits: Karine Monier, INMG-PGNM, Lyon 

Green-channel dSTORM imaging of AF488 fluorophore using Everspark 2.0 buffer

 AF488-coated beads were mounted in Everspark 1.0 or Everspark 2.0 buffer and imaged on a Vutara VXL (Bruker) before 3D dSTORM reconstruction.
Credits: Karine Monier, INMG-PGNM, Lyon 

Long-term imaging with Everspark 1.0

Left: Two centrosomes imaged the same Day (D0) and 7 days after mounting (D7) on the same slide stored in the dark at 4°C.  The typical 450 nm donut-like structure is visualised using a colour-coded scale encoded with the  IGOR software, where each point appears as a function of its localisation precision (5 to 60 nm; inverted rainbow colour scale). Labeling: Distal-appendages detected by immunofluorescence with AF647 in RPE-1 cells. 
Right: The number of blinking events per centrosome and the median of  the localization precision in nm are presented for each serie of 50,000 images recorded at D0 and D7 (left). 
Credits: Camille Fourneaux & Karine Monier, CRCL, Lyon

Donut-like structure of in-cellulo mature centrosome reconstructed after dSTORM imaging in Everspark 1.0 buffer

Each point is represented by its centroid (purple points) and its gaussian width (white). Intensity profil is displayed on the right with the measurement from peak to peak, in agreement with the size of the distal appendage crone. Labeling: distal-appendages detected by immunofluorescence with AF647 in RPE-1 cells.

Credits: Corentin Rousset, Karine Monier, CRCL Lyon

Snapshot analysis of SOX2 temporal dynamics after dSTORM imaging in Everspark 1.0 buffer

(a) 2D dSTORM image of active RNA polymerase II clusters (blue) and HaloTag-SOX2 10 clusters (colour map related to cluster distance). The colour map reflects the distance of SOX2 clusters from the nearest polymerase II cluster (green closest, and red farther). (b) A discreet subset of SOX2 clusters is within a 50 nm radius of the nearest RNA polymerase II cluster.

Credits: Bitong Li et al. (2024), Centenary Institute, University of Sydney

Publications about EverSpark:

Everspark technology has been initially developed by Karine Monier, Arnaud Favier and Christophe Place and published in Scientific Reports:
Provost, A., Rousset, C., Bourdon, L. et al. Innovative particle standards and long-lived imaging for 2D and 3D dSTORM. Sci Rep 9, 17967 (2019). https://doi.org/10.1038/s41598-019-53528-0
 

Discover three-color nanoscale imaging with Everspark 2.0 by watching this on-demand webinar by Karine Monier. 

Publications:

Dia2 formin controls receptor activity by organizing plasma membrane lipid partitioning at the nanoscale
Changting LI, Yannick HAMON, David MAZAUD, Pamela GONZALEZ TRONCOSO, Marie DESSARD, Hai-tao HE, Christophe LAMAZE, and Cedric M. BLOUIN. bioRxiv 2023.12.15.571857; doi: https://doi.org/10.1101/2023.12.15.571857

Nanoscale engagement and clusterization of Programmed death ligand 1 (PD-L1) in the membrane lipid rafts of Non-Small Cell Lung Cancer cells 
Martina Ruglioni, Simone Civita, Tiziano Salvadori, Sofia Cristiani, Vittoria Carnicelli, Serena Barachini,  Iacopo Petrini, Irene Nepita, Marco Castello, Alberto Diaspro, Paolo Bianchini, Barbara Storti, Ranieri Bizzarri,  Stefano Fogli and Romano Danesi   bioRxiv 2022.08.09.503318; doi: https://doi.org/10.1101/2022.08.09.503318

HIV-1 diverts actin debranching mechanisms for particle assembly and release in CD4 T lymphocytes
Rayane Dibsy, Erwan Bremaud, Johnson Mak, Cyril Favard, Delphine Muriaux
Nat Commun. 2023 Oct 31;14(1):6945. doi: 10.1038/s41467-023-41940-0  

Fluorescent Polymer-AS1411-Aptamer Probe for dSTORM Super-Resolution Imaging of Endogenous Nucleolin
Fabre L, Rousset C, Monier K, Da Cruz-Boisson F, Bouvet P, Charreyre MT, Delair T, Fleury E, Favier A. Biomacromolecules. 2022 May 12. doi: 10.1021/acs.biomac.1c01706. PMID: 35549176

Comparative analysis of ChAdOx1 nCoV-19 and Ad26.COV2.S SARS-CoV-2 vector vaccines.
Michalik S, Siegerist F, Palankar R, Franzke K, Schindler M, Reder A, Seifert U, Cammann C, Wesche J, Steil L, Hentschker C, Gesell-Salazar M, Reisinger E, Beer M, Endlich N, Greinacher A, Völker U. Haematologica. 2022 Apr 1;107(4):947-957. doi: 10.3324/haematol.2021.280154. PMID: 35045692

Metabolic biorthogonal labeling and dSTORM imaging of peptidoglycan synthesis in Streptococcus pneumoniae 
Jennyfer Trouve, Oleksandr Glushonkov and Cecile Morlot
Star Protocols, December 13, 2021. doi: 10.1016/j.xpro.2021.101006

Insights in ChAdOx1 nCoV-19 vaccine-induced immune thrombotic thrombocytopenia.
Greinacher A, Selleng K, Palankar R, Wesche J, Handtke S, Wolff M, Aurich K, Lalk M, Methling K, Völker U, Hentschker C, Michalik S, Steil L, Reder A, Schönborn L, Beer M, Franzke K, Büttner A, Fehse B, Stavrou EX, Rangaswamy C, Mailer RK, Englert H, Frye M, Thiele T, Kochanek S, Krutzke L, Siegerist F, Endlich N, Warkentin TE, Renné T. 
Blood. 2021 Dec 2;138(22):2256-2268. doi: 10.1182/blood.2021013231. PMID: 34587242

Superresolution Microscopy of Drosophila Indirect Flight Muscle Sarcomeres.
Szikora S, Novák T, Gajdos T, Erdélyi M, Mihály J. Bio Protoc. 2020 Jun 20;10(12):e3654. doi: 10.21769/BioProtoc.3654. eCollection 2020 Jun 20. PMID: 33659324

Identification of IQCH as a calmodulin-associated protein required for sperm motility in humans
Cavarocchi E, Sayou C, Lorès P, Cazin C, Stouvenel L, El Khouri E, Coutton C, Kherraf ZE, Patrat C, Govin J, Thierry-Mieg N, Whitfield M, Ray PF, Dulioust E, Touré A.  iScience. 2023 Jul 10;26(8):107354. doi: 10.1016/j.isci.2023.107354.

PGE2 inhibits TIL expansion by disrupting IL-2 signalling and mitochondrial function
Morotti M, Grimm AJ, Hope HC, Arnaud M, Desbuisson M, Rayroux N, Barras D, Masid M, Murgues B, Chap BS, Ongaro M, Rota IA, Ronet C, Minasyan A, Chiffelle J, Lacher SB, Bobisse S, Murgues C, Ghisoni E, Ouchen K, Bou Mjahed R, Benedetti F, Abdellaoui N, Turrini R, Gannon PO, Zaman K, Mathevet P, Lelievre L, Crespo I, Conrad M, Verdeil G, Kandalaft LE, Dagher J, Corria-Osorio J, Doucey MA, Ho PC, Harari A, Vannini N, Böttcher JP, Dangaj Laniti D, Coukos G. PGE2 inhibits TIL expansion by disrupting IL-2 signalling and mitochondrial function. Nature. 2024 May;629(8011):426-434. doi: 10.1038/s41586-024-07352-w. Epub 2024 Apr 24. PMID: 38658764; PMCID: PMC11078736.

Cis-B7:CD28 Interactions at Invaginated Synaptic Membranes Activate CD28 and Promote CD8+ T Cell Function and Anti-Tumor Immunity
Y. Zhao, C. Caron, Y.-Y. Chan, C. K. Lee, X. Xu,J.Zhang, T. Masubuchi, C. Wu, J.D. Bui, E. Hui. Cis-B7:CD28 Interactions at Invaginated Synaptic Membranes Activate CD28 and Promote CD8+ T Cell Function and Anti-Tumor Immunity. Immunity. 2023 June 13; 56(6): 1187–1203.e12. doi:10.1016/j.immuni.2023.04.005.

Anti-phase clustering of regulatory factors shapes gene bursting

Li B., Wong Y. Y,  Flores-Rodriguez N., Davidson T., Graus M. S., Smialkovska V.,  Ohishi H., Feldmann A., Ochiai H., Francois M.

bioRxiv 2024.09. 10.612363; doi: https://doi.org/10.1101/2024.09.10.612363