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A new Zeiss LSM 710 laser scanning microscope for the DCR LCI Core Facility

English title A new Zeiss LSM 710 laser scanning microscope for the DCR LCI Core Facility
Applicant Rothen-Rutishauser Barbara
Number 145003
Funding scheme R'EQUIP
Research institution Pneumologie (Erwachsene) Department for BioMedical Research Universität Bern
Institution of higher education University of Berne - BE
Main discipline Cellular Biology, Cytology
Start/End 01.12.2012 - 30.11.2013
Approved amount 275'835.00
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All Disciplines (4)

Discipline
Cellular Biology, Cytology
Experimental Cancer Research
Pathophysiology
Immunology, Immunopathology

Keywords (3)

Laser scanning microscopy; Live cell immaging; Cell biology

Lay Summary (German)

Lead
Das Departement Klinische Forschung (DKF) an der Universität Bern hat eine neue Live Cell Imaging (LCI) Core Facility aufgebaut. Diese ist bereits mit zwei Mikroskopen, welche optimal für die Untersuchung von Lebendzellkulturen mit Hellfeld- und Fluoreszenzmikroskopie sind, ausgestattet. Mit dem neuen konfokalen Laser Scanning Mikroskop, welches auf schnelle und sensitive 3D-Analyse von Fluoreszenz in lebenden Zell- und Gewebekulturen ausgerichtet ist, wird die Core Facility optimal ergänzt.
Lay summary

Inhalt und Ziel des Projektes

Das „Zeiss LSM 710“ bringt eine Reihe von essentiellen Eigenschaften und Neuerungen mit sich wie eine erhöhte Scangeschwindigkeit und Photoempfindlichkeit. Das Prinzip der konfokalen Mikroskopie erlaubt ausserdem die 3D-Lokalisierung und Charakterisierung von Proteinen und Partikeln innerhalb von einzelnen Zellen mit grosser Präzision. Das neue Beleuchtungs- und Detektionsdesign des „Zeiss LSM 710“ erlaubt die Möglichkeit der stufenlosen Spektraldetektion über den gesamten Wellenlängebereich bei gleichzeitiger Anwendung von bis zu 10 Farbstoffen. Ebenso gehört eine Inkubationskammer (Regulierung von Temperatur, Luftfeuchtigkeit und CO2 ) zur Ausstattung, welche das Durchfu¨hren von Live Experimenten über kürzere oder längere Zeit erlaubt.

Wissenschaftlicher und gesellschaftlicher Kontext des Projektes

Die LCI Core Facility mit dem neuen Konfokalen Mikroskop steht allen Forschungsgruppen am DKF sowie der Universität Bern zur Verfügung. Der Koordinator der Facility, Dr. Fabian Blank, sowie ein technischer Mitarbeiter, garantieren eine optimale Betreuung und Koordination der diversen Forschungsprojekte im Raum Bern. Zusätzlich wird die Facility für die Ausbildung von Masterstudenten und Doktorierenden benutzt.

Direct link to Lay Summary Last update: 19.11.2012

Responsible applicant and co-applicants

Publications

Publication
Interaction of biomedical nanoparticles with the pulmonary immune system.
Blank Fabian, Fytianos Kleanthis, Seydoux Emilie, Rodriguez-Lorenzo Laura, Petri-Fink Alke, von Garnier Christophe, Rothen-Rutishauser Barbara (2017), Interaction of biomedical nanoparticles with the pulmonary immune system., in Journal of nanobiotechnology, 15(1), 6-6.
Inhalation pathway as a promising portal of entry: What has to be considered in designing new nanomaterials for biomedical application?
Müller L, Lehmann AD, Johnston BD, Blank F, Wick P, Fink A, Rothen-Rutishauser B (2014), Inhalation pathway as a promising portal of entry: What has to be considered in designing new nanomaterials for biomedical application?, in Sahu SC Casciano DA (ed.), John Wiley & Sons, Ltd, West Sussex, UK, 205-221.
Quantification of nanoparticles at the single-cell level: an overview about state-of-the-art techniques and their limitations.
Vanhecke Dimitri, Rodriguez-Lorenzo Laura, Clift Martin J D, Blank Fabian, Petri-Fink Alke, Rothen-Rutishauser Barbara (2014), Quantification of nanoparticles at the single-cell level: an overview about state-of-the-art techniques and their limitations., in Nanomedicine (London, England), 9(12), 1885-900.
EphA2 signaling following endocytosis: role of Tiam1
Boissier P Chen J Huynh-Do U (2013), EphA2 signaling following endocytosis: role of Tiam1, in Traffic. , 14(12), 1255-1271.
Presence of the Gpr179nob5 allele in a C3H-derived transgenic mouse.
• Jasmin Balmer Rui Ji Thomas A. Ray Fabia Selber Max Gassmann Neal S. Peachey Ronald G. Gregg (2013), Presence of the Gpr179nob5 allele in a C3H-derived transgenic mouse., in Molecular Vision , 19, 2615-2625.
Size-dependent Uptake of Particles by Respiratory APC Populations and Trafficking to Regional Lymph Nodes
Blank F Stumbles PA Seydoux E Holt PG Fink A Rothen-Rutishauser B Strickland DH von Garnier C (2013), Size-dependent Uptake of Particles by Respiratory APC Populations and Trafficking to Regional Lymph Nodes, in Am J Respir Cell Mol Biol , 49(1), 67-77.
The guanine nucleotide exchange factor Tiam1: A Janus-faced molecule in cellular signaling
Boissier P Huynh-Do U (2013), The guanine nucleotide exchange factor Tiam1: A Janus-faced molecule in cellular signaling, in Cell Signal, 26(3), 483-491.
Inhalation pathway as a promising portal of entry: What has to be considered in designing new nanomaterials for biomedical application
• Müller L Lehmann AD Johnston BD Blank F Wick P Fink A Rothen-Rutishauser B, Inhalation pathway as a promising portal of entry: What has to be considered in designing new nanomaterials for biomedical application, in Sahu S Casciano DA (ed.), Wiley, UK.
Nanoparticles in the respiratory tract: Modulation of antigen-presenting cell function
Blank F Rothen-Rutishauser B von Garnier Ch, Nanoparticles in the respiratory tract: Modulation of antigen-presenting cell function, in J Environ Immunol Toxicol .

Collaboration

Group / person Country
Types of collaboration
Microscopy Imaging Centre Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Swiss Society of Optics and Microscopy (SSOM) Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results

Associated projects

Number Title Start Funding scheme
127586 Evolution of in vitro Intervertebral Disc Culture Systems: Two Degrees of Freedom Loading (Axial Compression and Torsion) to Study Region-Specific and Synergistic Degenerative Processes 01.01.2010 Project funding (Div. I-III)
141127 Alveolar-capillary microfluidic model for the analysis of lung injury, regeneration and repair 01.05.2012 Project funding (Div. I-III)
131266 Biomedical nanoparticles as immune-modulators 01.05.2011 NRP 64 Opportunities and Risks of Nanomaterials
125394 The link between aberrant MET signaling and DNA repair pathways in liver tumor cells as a target for sensitization to DNA damaging agents 01.01.2010 Project funding (Div. I-III)
123453 Clinical impact and pathophysiological mechanisms of rhinovirus infections in cystic fibrosis lung disease 01.05.2009 SNSF Professorships
122355 Pulmonary immune responses to biomedical nanoparticles and therapeutical applications 01.03.2009 Project funding (Div. I-III)
127628 Molecular mechanisms of granulocyte activation 01.11.2009 Project funding (Div. I-III)
130816 Natural killer cell function after liver resection: the role of extracellular ATP 01.04.2010 Project funding (Div. I-III)
140349 Exploring Peptide Topologies in Search for New Drugs 01.04.2012 Project funding (Div. I-III)
135565 Novel ways for improved cell replacement strategies in Parkinson’s disease: The potential of Nogo-A neutralization. 01.02.2012 Project funding (Div. I-III)
141102 Paracrine soluble factors released by induced pluripotent stem cells in lung regeneration and repair 01.11.2012 Project funding (Div. I-III)

Abstract

In the recent past, the concept of the core facility has proven to be a very efficient strategy to fulfill the demanding needs of a huge number of advanced research groups in one and the same place. The University of Bern has successfully established the Microscopy Imaging Centre (MIC), which is an inter-faculty organization. It involves many different institutions within the medical, veterinary medicine and natural sciences faculties. Through the MIC, expertise in the most up-to-date microscope technology is maintained at the University of Bern, continues to develop further and is accessible to all interested individuals and groups of which a large number belongs to the Medical Faculty. However, all the microscopes are still physically located with the individual research groups. In particular for the use of light microscopy imaging systems, the necessary commitment to this technology is often difficult to maintain by individual laboratories, especially when optical imaging is not a major focus for the laboratory.The Department of Clinical Research (DCR) establishes and runs core facilities for researchers of the Inselspital and the Medical Faculty. Since November 2010 the new research building of the DCR at Murtenstrasse 50 (Mu50), which was financed by the Inselstiftung, is hosting many different research groups from the DCR and the ARTORG Centre. For the new DCR Live Cell Imaging (LCI) Core Facility rooms in the basement of the building were prepared to host different microscopes for live cell imaging. This Facility is headed by Dr. Fabian Blank since March 2012, and aims to provide equipment and expertise necessary to facilitate the use of microscopy in research within the range of sample preparation, imaging and data processing, restoration and storage. Lectures and practical courses within the MIC activities in those fields are provided on a regular basis. Currently the LCI Core Facility has a Leica “DMI4000 B” fluorescence microscope for conventional state-of-the-art fluorescence and bright field imaging techniques for the investigation of living and fixed cell- and tissue cultures. In addition, the facility includes a “Nikon Biostation CT”, which provides a fully automated long term monitoring of living cells and tissues using a phase contrast and fluorescence microscope integrated inside an incubator. All these instruments were chosen to optimally complement each other.However, the LCI Core Facility, as it is equipped today, is lacking a confocal laser scanning device optimized for fast and sensitive 3D and time-lapse 3D (4D) analyses of fluorescence inside living cells and tissues. There is a steadily growing demand from research groups for such a microscope. Since the confocal microscopes within the MIC are localized in other institutes within the university campus and are already used to their full potential, increased use, especially for living cell experiments, is not possible. In addition, the newest microscope in Bern was purchased in 2007 and does no longer fulfill the requirements for scanning speed and sensitivity for long term observations. The DCR has therefore decided to purchase a new “Zeiss LSM 710” laser scanning microscope to complete and optimize the LCI core facility. This device features a number of essential as well as novel properties and improvements: for example the scanning speed and the photosensitivity have been extensively improved, which leads to strongly reduced phototoxicity. As a consequence, with this instrument it is possible to scan living samples more often and for a longer time period at maximal resolution without any adverse effects for the cells. On the campus of the University of Bern this microscope will be the fastest point scanning system for living cells without loss in resolution. This fully equipped system consists of the confocal microscope itself, 4 different filter systems for visual work, 7 different objectives optimized for live fluorescence imaging including long-distance objectives, 3 different spectral fluorescence channels with the laser lines 405, 458, 488, 514, 561, and 633, a temperature and CO2 controlled incubation chamber, an automatized scanning table, a high resolution camera and a complete workstation including acquisition software. Since the number of cell-/tissue-based experiments is increasing in most labs, the acquisition of the Zeiss LSM 710 laser scanning microscope is an urgent need for researchers at the University of Bern. Furthermore in order to stay competitive, for clinical research groups on the campus of the University of Bern it is an essential prerequisite to have the best available technology provided. This microscope would perfectly complete the two other microscopes already located in the DCR LCI core facility by offering the possibility to investigate biological samples at higher resolution in 2D, 3D or even 4D. All instruments within the LCI core facility will be openly accessible within the MIC for all researchers on the Insel campus.
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