Bioimaging Unit

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The HPI Bioimaging Unit (HPI-BU) is one of the most modern and well-equipped Imaging Facilities in Greece and a member of the Greek Bioimaging-GR Consortium and the European Light Microscopy Initiative (ELMI- www.embl.org/elmi/). HPI-BU is an open-access platform aiming to provide advanced Light microscopy and image analysis services to HPI, national or international academic scientists.

The HPI-BU has created an international wide network with top imaging groups in the EU and the Institut Pasteur Paris that have largely contributed to the development and establishment of cutting-edge technologies in this Facility. European and National grants such as BIOIMAGING-GR – Ministry of Education Infrastructure Grant (2017-2020), Stavros Niarchos Foundation Grant: (2016-2020), KRHPIS I-II (2015-2018), NEUROSIGN EU FP7 Program (2010-2013) have supported the significant upgrading of the HPI-BU which has become a hub of a 2-photon Intravital Imaging system in the Bioimaging-GR Consortium, as well as fast live cell imaging systems.

Τhe main state-of-the art methodologies currently in place in the HPI-BU are: a) 2- photon intravital imaging in the CNS of transgenic mice, b) short and long time live cell imaging and c) bioimage analysis using commercially available and open-access image analysis software.

The HPI-BU has emerged within the Institut Pasteur International Network as a node for advanced imaging training and scientific collaborations. The unit has organized many training activities and several publications have been published in peer review journals with the participation or acknowledgement of the HPI-BU.

Our strategic goal is to develop the appropriate tools and technology that will allow us to monitor in real time disease progression in a wide range of communicable and non-communicable disease models, to develop live cell Calcium imaging, as well as multiple parameter middle-throughput and high-throughput screening, aiming to the understanding of biological processes and screening for novel therapeutics.

Registration

New users of the Bioimaging Unit (BU) of HPI, as well as current users, who are starting a new project, should submit a project description in a form provided at the HPI-BU. We need all necessary information to organize on time introductory sessions for using the instruments, as well as for teaching and for support in sample preparation techniques and new imaging protocols.

External users

The HPI’s imaging systems can be used by external users, who should contact the responsible scientists of the Unit in order to arrange the exact time of the microscope booking.

  • ImageJ/Fiji macro workshop. 6-10 May 2019

A 5-day hands-on workshop was organized in the Hellenic Pasteur Institute in Athens Greece in collaboration with the Image Analysis hub of the Institut Pasteur Paris with the objective to provide theoretical and hands-on training in image analysis and automation tools.

The workshop programme included Lectures and hand-on sessions on: the basics of ImageJ/Fiji, object detection and segmentation, machine learning with the use of WEKA and Ilastik, single particle tracking with Trackmate, ImageJ Macro programming language, batch processing using macros and advanced macro programming in Fiji, while the last day of the workshop was dedicated to help the trainees analyze their own data and apply the new knowledge they gained.

The trainers were Dmitry Ershov and Sébastien Herbert, who kindly accepted our invitation to train the members of our Unit and external scientists on the ImageJ Macro programming language and machine-learning software. The trainees were 5 scientists from the Hellenic Pasteur institute (4 of which were members of the Bioimaging-GR program) and 3 external scientists from the University and other research centers in Athens.

Sponsors:

BIOIMAGING-GR, Hellenic Pasteur Institute

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  • «REVEALING LIFE’S HIDDEN SECRETS THROUGH NEW TECHNOLOGY» (5-6/06/2018) in collaboration with Leica Microsystems

 

The 2-day workshop was organized at the Hellenic Pasteur Institute in Athens, with the support of Leica Microsystems, the Bioimaging-GR program and the Hellenic Pasteur Institute. The objective of the workshop was to provide theoretical and practical training in advanced applications of microscopy in life sciences with an emphasis on 2-photon microscopy and confocal fluorescence microscopy through:

 

  • Lectures by Leica Microsystems instructors and Greek scientists
  • Practical training in the use of confocal microscopes
  • Discussions with experts in the field of microscopy

 

35 trainees followed the theoretical and practical part, while a significant number of participants only attended the theoretical part of the seminar.

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  • RIIP International course “Cell biology and Infection: Digital Image processing/Analysis tools for quantitative light microscopy imaging” (4-8 July 2016)

A 5-day practical course was organized by the Hellenic Pasteur Institute in Athens Greece during 4-8 July 2016 with objective to provide theoretical and practical training in basic and advanced concepts and methods of Bioimage informatics. The course Program included:

  • Lectures focused on concepts and methodology for Processing/Analysis of Light Microscopy Digital data (images and videos) and selected topics of Cell Biology and Biology of Pathogen Host interaction.
  • Computer practical sessions on the quantitative analysis of microscopy image datasets using the open source software Icy and ImageJ/Fiji
  • Informal discussions and tutorials with experts in the field

 

  • European FP7 Regpot-2010 ‘Neurosign’ theoretical and practical workshop “Live Cell Imaging and Electrophysiology” (1-4/10/2013)

The workshop was conducted during 1-4 October 2013 in the Hellenic Pasteur Institute in Athens Greece. The aim of this workshop was to provide post graduate and post-doctoral scientists with a conceptual and practical understanding of CNS live cell imaging in whole animal level, as well as electrophysiology. During the practicals, the 20 selected participants had the chance to observe both live and video recorded experiments including surgical procedures for creating brain and spinal cord windows, intravital Multiphoton imaging and whole-cell and single-channel recordings, using the patch-clamp technique applied on mammalian cells. The funding was provided by the FP7 REGPOT 2010-1 EC Program “NeuroSign” that aims at the establishment of a Centre of Excellence for the study of Neurosignalling during nervous system function and dysfunction at the Hellenic Pasteur Institute.

  • RIIP Regional CourseDigital image processing/ analysis tools in Light Microscopy: From the basics and beyond” (10-17/6/ 2013)

An 8-day course was organized in the Hellenic Pasteur Institute in Athens, Greece during 10-17 June 2013. The course objective was to provide theoretical and practical training in basic and advanced concepts and methods of digital image analysis and in the use of several open source and commercial software for quantitative assessment of fluorescence microscopy data through:

    1. Lectures
    2. Hands – on practical training sessions with participants’ microscopy data or data provided by the instructors.
    3. Informal discussions and tutorials with experts in the field.
    4. Practical sessions on the following image analysis software: Imaris, Scientific Volume Imaging (SVI), Image J/Fiji, Icy bioimage analysis, Cell profiler and Timm’s Tracking Tool.

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PUBLICATIONS 2018 - from projects carried out in HPI-BU

Senescent cells in Giant Cell Arteritis display inflammatory phenotype participating in tissue injury via IL-6 dependent pathways.

Veroutis D, Argyropoulou OD, Goules AV, Kambas K, Palamidas DA, Evangelou K, Havaki S, Polyzou A, Xingi E, Karatza E, Boki K, Cavazza A, Kittas C, Thanos D, Ricordi C, Marvisi C, Muratore F, Galli E, Croci S, Salvarani C, Gorgoulis VG, Tzioufas AG.

Annals of the Rheumatic Diseases 2024 Feb 15:ard-2023-224467.

PMID: 38050005

 

Characterization of the First Secreted Sorting Nexin Identified in the Leishmania Protists.   Tziouvara O.  1 Petsana M. , KourounisD., Papadaki A.,  Basdra E.,  Braliou G. G, BoletiH.          Int J Mol Sci. 2024 Apr 7;25(7):4095.

PMID: 38612903

 

LPS-Induced Systemic Inflammation Affects the Dynamic Interactions of Astrocytes and Microglia with the Vasculature of the Mouse Brain Cortex. Xingi E, Koutsoudaki P, Thanou I, Phan M.S, Margariti M, Scheller A, Tinevez J-Y, Kirchhoff F, Thomaidou D. Cells (2023) May 17;12(10):1418.

PMID: 37408252

 

A miR-124-mediated post-transcriptional mechanism controlling the cell fate switch of astrocytes to induced neurons. Papadimitriou E,  Koutsoudaki P.N, Thanou IKaragkouni D, Karamitros T, Chroni-Tzartou D, Gaitanou M, Gkemisis C, Margariti M, Xingi ETzartos SJ, Hatzigeorgiou AG, Thomaidou D

Stem Cell Reports 2023 Mar 13;S2213-6711(23)00056-5.

PMID: 36963393

 

Down-regulation of KLF2 in lung fibroblasts is linked with COVID-19 immunofibrosis and restored by combined inhibition of NETs, JAK-1/2 and IL-6 signalingChrysanthopoulou A, Antoniadou C  Natsi A-M , Gavriilidis E  Papadopoulos V, Xingi EDidaskalou S, Mikroulis D Tsironidou V, Kambas K Koffa M, Skendros P, Ritis K

 Clin Immunol 2023 Feb;247:109240.

PMID: 36693535

 

Human monoclonal natural IgG antibodies can penetrate MDA-MB-231 cells and transport intracellularly paclitaxel-loaded gold nanorods. Stivarou S, Papaioannou L, Sarrigeorgiou I, Avgoustakis K, Lymberi P.

Journal of Drug Delivery Science and Technology Vol. 80, February 2023, 104109

https://doi.org/10.1016/j.jddst.2022.104109

 

New Affordable Methods for Large-Scale Isolation of Major Olive Secoiridoids and Systematic Comparative Study of Their Antiproliferative/Cytotoxic Effect on Multiple Cancer Cell Lines of Different Cancer Origins. Papakonstantinou A, Koumarianou P, Rigakou A, Diamantakos P,  Frakolaki E, Vassilaki N, Chavdoula E, Melliou E, Magiatis P, Boleti H

Int J Mol Sci 2022 Dec 20;24(1):3.

PMID: 36613449

 

Neutrophil extracellular traps in giant cell arteritis biopsies: presentation, localization and co-expression with inflammatory cytokines.  Palamidas DA,  Argyropoulou OD, Georgantzoglou N Karatza E, Xingi EKapsogeorgou EK,  Anagnostopoulos CD,  Lazaris AC, Ritis K .

Rheumatology (Oxford) 2022 Apr 11;61(4):1639-1644.

PMID: 34260696

 

Exploiting the Role of Hypoxia-Inducible Factor 1 and Pseudohypoxia in the Myelodysplastic Syndrome Pathophysiology
Stergiou IE, Kambas K, Poulaki A, Giannouli S, Katsila T, Dimitrakopoulou A, Vidali V, Mouchtouris V, Kloukina I, Xingi E, Pagakis SN, Probert L, Patrinos GP, Ritis K, Tzioufas AG and Voulgarelis M.

Ιnt. J. Mol. Sci. 2021, 22, 4099.

PMID: 33921064

 

The Leishmania donovani LDBPK_220120.1 Gene Encodes for an Atypical Dual Specificity Lipid- Like Phosphatase Expressed in Promastigotes and Amastigotes;Substrate Specificity, Intracellular Localizations, and Putative Role(s).

Papadaki A, Tziouvara O, Kotopouli A, Koumarianou P, Doukas A, Rios P, Tardieux I, Köhn M
and Boleti H

Front Cell Infect Microbiol 2021 Mar 25;11:591868. doi: 10.3389/fcimb.2021.591868.

PMID: 33842381

 

Novel cell-based analysis reveals region-dependent changes in microglial dynamics in grey matter in a cuprizone model of demyelination.  Roufagalas I, Avloniti M, Fortosi A, Xingi E, Thomaidou D, Probert L, Kyrargyri V.

Neurobiology of Disease, 2021 Sep;157:105449. doi: 10.1016/j.nbd.2021.105449.

PMID: 34274460

 

Total Phenolic Fraction (TPF) from Extra Virgin Olive Oil: Induction of apoptotic-like cell death in Leishmania spp. promastigotes and in vivo potential of therapeutic immunomodulation. Karampetsou K, Koutsonι OS, Gogou G, AAngelis A, Skaltsounis LA, Dotsika E. PLoS Negl Trop Dis 15(1) 2021: e0008968.

PMID: 33428610

 

High Content Screening and Proteomic Analysis Identify a Kinase Inhibitor that rescues pathological phenotypes in a Patient-Derived Model of Parkinson's Disease. Antoniou N, Prodromidou K, Kouroupi G, Samiotaki M, Panayotou G, Xilouri M, Stefanis L, Grailhe R, Taoufik E,  Matsas R. Bioarchives 2021, doi: https://doi.org/10.1101/2020.06.12.148031

 

Increased autophagy/mitophagy levels in primary tumours of patients with pancreatic neuroendocrine neoplasms.  

Daskalakis K, Alexandraki KI, Kloukina I, Kassi E, Felekouras E, Xingi E, Pagakis SN, Tsolakis AV, Andreakos E, Kaltsas G, Kambas K.

Endocrine 2020 May;68(2):438-447. doi: 10.1007/s12020-020-02228-1.

PMID: 32114655

 

Leishmania dual-specificity tyrosine-regulated kinase 1 (DYRK1) is required for sustaining Leishmania  stationary phase phenotype

Rocha VPC, Dacher M,  Young SA, Kolokousi F, Efstathiou A,  Späth GF,  Soares MB, Smirlis D.

Mol Microbiol. 2020 May;113(5):983-1002. doi: 10.1111/mmi.14464.

PMID: 31975452

 

Combining RANK/RANKL and ERBB-2 targeting as a novel strategy in ERBB-2-positive breast carcinomas. Zoi I, Karamouzis MV, Xingi E, Sarantis P, Thomaidou D, Lembessis P, Theocharis S, Papavassiliou AG.

Breast Cancer Res. 2019 Dec 3;21(1):132. doi: 10.1186/s13058-019-1226-9.

PMID:31796128

 

Estrogen receptor beta increases sensitivity to enzalutamide in androgen receptor-positive triple-negative breast cancer.

Anestis A, Sarantis P, Theocharis S, Zoi I, Tryfonopoulos D, Korogiannos A, Koumarianou A, Xingi E, Thomaidou D, Kontos M, Papavassiliou AG, Karamouzis MV.                                                                    

J Cancer Res Clin Oncol. 2019 May;145(5):1221-1233. doi: 10.1007/s00432-019-02872-9. PMID:30805773

 

Heterologous expression of the mammalian sodium-nucleobase transporter SNBT1 in Leishmania tarentolae.

Doukas A, Karena E, Botou , Papakostas K, Papadaki A, Tziouvara O, Xingi E, Frillingos S, Boleti H.

Biochim Biophys Acta Biomembr. (2019). 1861(9):1546-1557. doi:10.1016/j.bbamem.2019.07.001. 

PMID:31283918

 

CHUK/IKKalpha loss in lung epithelial cells enhances NSCLC growth associated with HIF up-regulation.

Chavdoula E., Habiel D.M., Roupakia E., Markopoulos G.S., Vasilaki E., Kokkalis A., Polyzos A.P, Boleti H., Thanos D., Klinakis A., Kolettas E., Marcu K.B.

 Life Science Aliance, (2019). 2(6). http://doi.org/10.26508/lsa.201900460

 

Emerging Role of l-Dopa Decarboxylase in Flaviviridae Virus Infections.

Frakolaki E, Kalliampakou KI, Kaimou P, Moraiti M, Kolaitis N, Boleti H, Koskinas J, Vassilacopoulou D, Vassilaki N.

Cells 8(8). (2019) pii: E837. doi: 10.3390/cells8080837.

PMID:31387309

 

Increased Anxiety-Related Behavior, Impaired Cognitive Function and Cellular Alterations in the Brain of Cend1-deficient Mice.

Segklia K, Stamatakis A, Stylianopoulou F, Lavdas AA, Matsas R.

Front Cell Neurosci. 2019 Jan 29;12:497. doi: 10.3389/fncel.2018.00497.

 PMID: 30760981

 

In Vivo Phenotyping of Familial Parkinson’s Disease with Human  Induced Pluripotent Stem Cells: A Proof-of-Concept Study
Zygogianni O, Antoniou N, Kalomoiri M, Kouroupi G, Taoufik E, Matsas R
Vol.:(0123456789)Neurochemical Research (2019) 44:1475–1493
https://doi.org/10.1007/s11064-019-02781-w.

PMID: 30989481

 

Systemic activation of NLRP3 inflammasome in patients with severe primary Sjögren’s syndrome fueled by inflammagenic DNA accumulations.  (2018). Vakrakou AG, Boiu S, Ziakas PD, Xingi E, Boleti H, Manoussakis MN. J Autoimmun. Jul; 91:23-33. PMID: 29551295

 

The Scientists supporting the HPI-BU have participated either as members or as Scientific responsible persons in several grant applications succeeding to attract funding for the Facility either for organizing courses or for upgrading the equipment and development of new methodology from the following grants:

  •  International Network of Pasteur Institutes (IPIN) PTR Grant (2020-2022): “Microglia Imaging in Alzheimer’s Disease”. Scientific Responsibles:  Dr. D. Thomaidou, Dr. M. Costa (Institut Pasteur Lille), Dr. J-Y. Tinevez, Institut Pasteur Paris.
  •  ΒΙΟΙΜAGING GR 2018-2020: “Hellenic Research Infrastructure for the Imaging and Observation of Fundamental Processes in Biology and Medicine” (MIS 5002755) which is implemented under the Action “Reinforcement of the Research and Innovation Infrastructure”, funded by the Operational Programme “Competitiveness, Entrepreneurship and Innovation” (NSRF 2014-2020) and co-financed by Greece and the European Union (European Regional Development Fund, coordinator Dr. Tavernarakis, IMBB Forth).
  •  Stavros Niarchos Foundations Sponsorships 2016-2020 “Development of innovative Biological products and health services for infectious and neurodegenerative diseases” (Coordinators Drs. R. Matsa and V. Myriagou)
  •  Ηellenic State Scholarship Foundation (Ι.Κ.Υ.) and German Burau for Academics’ Exchanges (DAAD): IKYDA program 2014-2015: Astroglia activation and reprogramming in brain injury and repair: analysis by in vivo imaging (2P-LSM) of the brain (Scientific Responsible Dr. D. Thomaidou).
  •  RIIP (Institut Pasteur International Network) International course 2016:
    ” Cell Biology and infection: Digital Image Processing/Analysis Tools for Quantitative Light Microscopy Imaging”. Scientific Responsible Dr. H. Boleti
  •  FP7 EU program 264083 NEUROSIGN 2010-2013: Development of Centre of Excellence in Neurosciences (coordinators. Dr.S. Tzartos– Dr. R. Matsa & Dr. L. Probert (Dr. D. Thomaidou Responsible for WP of purchasing the Infrastructure equipment for in vivo imaging).
  •  RIIP (Institut Pasteur International Network) Regional course 2013:
    “Digital image processing/analysis tools in Light Microscopy: From the basics and beyond”. Scientific Responsible Dr. H. Boleti
  •  Competitive grant from the Greek General Secretariat of Research and Technology in the context οf European program “Human Networks for training in Research and Technology” 2004-2006 –Establishment of the Greek Light Microscopy Network Title: “Applications of Light Microscopy in Biomedical research and Diagnosis” In collaboration with the U. of Ioannina & U. of Crete Coordinator of the project and of the Network, HPI-LMU. (Scientific Responsible ¨Drs H. Boleti & D. Thomaidou)
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