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Imaging of Deep-Lying Tissues with Enhanced Resolution and Sensitivity to Study Plant Development

English title Imaging of Deep-Lying Tissues with Enhanced Resolution and Sensitivity to Study Plant Development
Applicant Vermeer Joop
Number 164086
Funding scheme R'EQUIP
Research institution Département de Biologie moléculaire des plantes Université de Lausanne
Institution of higher education University of Zurich - ZH
Main discipline Cellular Biology, Cytology
Start/End 01.12.2015 - 30.11.2016
Approved amount 490'000.00
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All Disciplines (3)

Discipline
Cellular Biology, Cytology
Botany
Embryology, Developmental Biology

Keywords (11)

mechanical constraints; pollen tube; plant hormones; plant reproduction; mycorrhiza; disease resistance; cell biology; deep tissue live imaging; chromatin; gametophyte; plant development

Lay Summary (German)

Lead
Das neue, für viele unterschiedliche Anwendungen ausgelegte Mikroskop erlaubt tiefer gelegene Gewebeschichten mit einer erhöhten Auflösung und Empfindlichkeit zu beobachten. Damit können an der Universität Zürich Entwicklungsprozesse der Pflanze auf einer völlig neuen Ebene verfolgt werden.
Lay summary

Die Beobachtung von lebenden Zellen ist eine Voraussetzung um entwicklungsabhängige Prozesse zu studieren. Es hat sich gezeigt, dass die konfokale Mikroskopie eine überragende Rolle für die Darstellung von subzellulären Prozessen bei lebenden Zellen spielt und somit unverzichtbar ist, um offene Fragen zu Entwicklungsprozessen zu beantworten. Verglichen mit früheren Mikroskopen erlaubt das neue konfokale Mikroskop eine bessere und effizientere Beobachtung der Proben. Dabei können mit diesem Mikroskop auch grössere und dickere Proben mit einer höheren Auflösung und Empfindlichkeit beobachtet werden. Dieses sogenannte multi-photon Mikroskop mit dem neuen, revolutionären Airyscan Detektor wird den Wissenschaftlern an der Universität Zürich neue, bisher ungeahnte Felder der (Pflanzen)biologie eröffnen und so erlauben in neue Domänen vorzustossen. Zusätzlich, und für Pflanzenwissenschaftler äusserst wichtig, wird es das neue, flexible Detektionssystem erlauben Signale von fluoreszierenden Proteinen und der pflanzlichen Autofluoreszenz besser zu unterscheiden und damit auch Fehlinterpretationen zu verhindern.

 

Direct link to Lay Summary Last update: 20.11.2015

Responsible applicant and co-applicants

Publications

Publication
Root branching toward water involves posttranslational modification of transcription factor ARF7
Orosa-Puente Beatriz, Leftley Nicola, von Wangenheim Daniel, Banda Jason, Srivastava Anjil K., Hill Kristine, Truskina Jekaterina, Bhosale Rahul, Morris Emily, Srivastava Moumita, Kümpers Britta, Goh Tatsuaki, Fukaki Hidehiro, Vermeer Joop E. M., Vernoux Teva, Dinneny José R., French Andrew P., Bishopp Anthony, Sadanandom Ari, Bennett Malcolm J. (2018), Root branching toward water involves posttranslational modification of transcription factor ARF7, in Science, 362(6421), 1407-1410.
Diffusible repression of cytokinin signalling produces endodermal symmetry and passage cells
Andersen Tonni Grube, Naseer Sadaf, Ursache Robertas, Wybouw Brecht, Smet Wouter, De Rybel Bert, Vermeer Joop E. M., Geldner Niko (2018), Diffusible repression of cytokinin signalling produces endodermal symmetry and passage cells, in Nature, 555(7697), 529-533.
Green light for quantitative live-cell imaging in plants
Grossmann Guido, Krebs Melanie, Maizel Alexis, Stahl Yvonne, Vermeer Joop E. M., Ott Thomas (2018), Green light for quantitative live-cell imaging in plants, in Journal of Cell Science, 131(2), jcs209270-jcs209270.
In Vivo Imaging of Diacylglycerol at the Cytoplasmic Leaflet of Plant Membranes
Vermeer Joop E.M., van Wijk Ringo, Goedhart Joachim, Geldner Niko, Chory Joanne, Gadella Theodorus W.J., Munnik Teun (2017), In Vivo Imaging of Diacylglycerol at the Cytoplasmic Leaflet of Plant Membranes, in Plant and Cell Physiology, 58(7), 1196-1207.

Collaboration

Group / person Country
Types of collaboration
Eva Zažímalová Czech Republic (Europe)
- in-depth/constructive exchanges on approaches, methods or results
John Bowman / Monash University Australia (Oceania)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Alexis Maizel / COS Heidelberg Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Karin Ljung / Umea University Sweden (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Malcolm Bennett / CPIB Nottingham Great Britain and Northern Ireland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
H. Bouwmeester / WUR Netherlands (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Luca Comai / UC Davis United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
- Industry/business/other use-inspired collaboration
Jiri Friml / IST Austria (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Niko Geldner / University of Lausanne Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Dupont Pioneer / USA United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
- Industry/business/other use-inspired collaboration
Jean Philippe Vielle Calzada / Langebio Cinvestav Mexico (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
- Industry/business/other use-inspired collaboration
Eva Benkova / IST Austria (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Y. Lee /University of Pohang Korean Republic (South Korea) (Asia)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Milena Ouzunova / KWS Saat AG Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Industry/business/other use-inspired collaboration
B. Halkier Denmark (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Andreas Houben / IPK Gatersleben Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
- Industry/business/other use-inspired collaboration
Evans Lagudah / CSIRO Agriculture Flagship Canberra Australia (Oceania)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Peggy Ozias-Akins / University of Georgia Research Foundation United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
- Industry/business/other use-inspired collaboration
Tom Beeckman / VIB Belgium (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Anne Koltunow / CSIRO Australia (Oceania)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
- Industry/business/other use-inspired collaboration
Marc Albertsen and Shai Lawit / DuPont Pioneer United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
- Industry/business/other use-inspired collaboration

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Auxins and Cytokinins in Plant Development 2018 Poster Shaping signaling landscapes during organ development 01.07.2018 Prague, Czech Republic Vermeer Joop;
ICAR2018 Talk given at a conference Functional deep imaging using 2-photon microscopy 25.06.2018 Turku, Finland Vermeer Joop;
FASEB Mechanisms in Plant Development Talk given at a conference Cell volume regulation during lateral root formation 30.07.2017 Saxtons River, United States of America Vermeer Joop;
Auxin gathering in Brno Talk given at a conference Cell volume regulation during lateral root formation in Arabidopsis 08.12.2016 Brno, Czech Republic Vermeer Joop;
Auxin2016 Talk given at a conference Cell volume regulation during lateral root formation in Arabidopsis 20.10.2016 Hainan, China Vermeer Joop;


Knowledge transfer events



Self-organised

Title Date Place
Advanced course on 3D microscopy imaging of plant tissues and image processing 07.12.2016 Zurich, Switzerland

Associated projects

Number Title Start Funding scheme
154694 Functional characterization of broad-spectrum disease resistance in cereals 01.01.2015 Ambizione
149974 Mechanisms and role of linker histones dynamics in plant reproduction 01.01.2014 Project funding
149459 Defining Specificity in Cytokinin Ligand Localization and Signaling Interpretation 01.10.2013 Project funding
144081 Molecular interactions of wheat and fungal pathogens: Lr34-based, durable resistance and the wheat-powdery mildew pathosystem 01.01.2013 Project funding
147074 Transcriptional and post-translational regulation of Arabidopsis b-amylase 1 during drought stress 01.05.2013 Project funding
157524 Talking with the neighbours: Understanding spatial accommodation during plant development 01.10.2015 SNSF Professorships
152831 Investigation of phytohormone transport and the fate of hypodermal passage cells, a cell type specialized to release strigolactones 01.05.2014 Project funding
141245 The Genetic and Molecular Basis of Gametogenesis and Maternal Effects in Arabidopsis 01.05.2012 Project funding

Abstract

In recent years, fluorescence-based microscopical imaging of living tissue has greatly advanced our understanding of plant developmental processes at the cellular level. This has largely been possible due to the use of Arabidopsis as a model species. The fast and straightforward generation of stable transgenic lines expressing fluorescent cellular markers allows for 3-dimensional (3D) imaging of developmental processes using confocal laser scanning microscopy (CSLM). The intimate relationship between the organization of tissues in all cell layers, their functional specialization, and the overall tissue morphology implies that a comprehensive understanding can be only be achieved using non-invasive imaging approaches throughout the depth of a tissue. Yet, 3D-microscopical imaging of living plant tissue is confronted by specific challenges including, (i) the inherent lack of optical clarity and strong light-scattering nature of plant cells [due to cell walls and numerous autofluorescent compounds] that prevents an efficient penetration/readout of the excitation/emitted light in deep tissue layers; (ii) the relative thickness of plant tissues exceeding the working distance of most objective lenses; and (iii) the moderate to low output signals from spatially and temporally restricted fluorescent markers are poorly captured with standard detectors. These technical limitations severely impede several research projects at the Institute of Plant Biology (IPB) that focus on cell-based mechanical constraints in organogenesis, hormone signaling in physiological responses, cellular dynamics in reproduction and pathogen responses in several plant species e.g. Arabidopsis, Marchantia, petunia, maize and wheat. All these research projects rely on the analysis of structural, physiological and signaling components in living, intact plant tissues throughout multiple cell layers and at subcellular resolution. Novel, non-invasive imaging techniques that offer fast imaging in deep tissue layers with enhanced resolution and sensitivity can overcome these limitations. To make a quantum leap in our capacity to analyze developmental processes at subcellular resolution and in order to remain competitive with top plant research groups in Europe we are applying for an all-in-one confocal/multiphoton laser scanning microscopy system that offers, in addition to the traditional confocal mode, (i) multiphoton excitation for imaging of deep-lying tissues, and (ii) an innovative Airyscan detector that combines high sensitivity with enhanced resolution properties. These requirements are currently met by a unique, commercially available system: the Zeiss LSM880. This confocal system can be equipped with near infrared lasers for multiphoton excitation and the revolutionary Airyscan detector (a 32 GaAsP-array detector offering the best currently available sensitivity], providing a 1.7x increase in resolution in all dimensions resulting in a 5x smaller confocal volume, without limitations regarding sample depth and acquisition speed. In addition, the multiphoton laser also allows to specifically ablate cells in deep-lying tissues. These ablation experiments are essential to our projects aiming at manipulating mechanical constraints and to assess the role of the positional information and interactions with surrounding cells during development. We, as well as Urs Ziegler (head of the Center for Microscopy and Image Analysis, ZMB) have tested the LSM880 system with Airyscan and multiphoton excitation (April 2015) and agree that this is currently the best system to solve the above problems. Yet, as other systems with similar properties are expected to become available, we will thoroughly and objectively assess them to choose the optimal system for our research projects.The high-end technology offered by the proposed all-in-one multiphoton confocal imaging system will provide a major impulse for our studies in multiple species and will help us to perform internationally competitive, top-level research at IPB. It will uniquely complement our existing high-end microscopy imaging platform comprising wide-field fluorescence microscopes and CSLM instruments, thereby considerably increasing the attractiveness and international competitiveness of IPB. Importantly, the ZMB of the University of Zürich does not possess the imaging instrument as requested here. In his continuous efforts to foster excellence in biological imaging, and with the shared vision that this instrument will meet interest beyond the plant science community, the Head of ZMB, Urs Ziegler, fully supports our proposal. Consequently, the requested equipment, while physically located at IPB, will be imbedded in the ZMB infrastructure thereby offering access to all ZMB users, thus guaranteeing optimal maintenance and user training. In the following part, we will describe several projects for which the requested equipment will be of central importance.
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