This Summer school was organized by the Saclay Plant Sciences (SPS) network, one of the largest European plant sciences communities and was hosted by the cytology and imaging platform of the Institut Jean Pierre Bourgin - Plant Sciences in Versailles.

Microscopy is a fundamental tool for understanding the functioning of plants at the cellular to molecular scale. Recent technological advances (super-resolution, fluorescence lifetime imaging, biosensors...) now make it possible to address new scientific questions. This Summer School provided theoretical and practical insights into these aspects for 19 outstanding and enthusiastic PhD students or young postdoctoral researchers divided in small groups. Participants had the opportunity to discuss the advantages and disadvantages of different modalities, sample preparation, image acquisition, and data processing throughout the week.

Program and speakers

Download the program of the Summer School

The Summer School included:

> A set of theoretical lectures (~9 hours) on the major issues in advanced imaging: nanoscopy (super resolution), fluorescence lifetime analysis, ratiometry analysis, their uses, advantages or limitations, data processing, etc. The lectures were given by experts in the field, who were available for discussions during the Summer School, giving the participants an insight into the latest research findings and identifying key open questions in the field.

Speakers:
Arnaud Gautier (Sorbonne University, Paris, France)
Joachim Goedhart (University of Amsterdam, The Netherlands)
Julien Gronnier (Technical University of Munich, Germany)
Kalina Haas (Jean-Pierre Bourgin Institute - Plant Sciences, Versailles, France)
Eric Hosy (Interdisciplinary Institute for NeuroScience, Bordeaux, France)

> A set of practical sessions (~24 hours + Restitution) to tackle real-life approaches on plant samples and compare different modalities of conventional confocal microscopy, super-resolution and fluorescence lifetime analysis. All Summer School participants had access to all practical sessions, in groups of 3 or 4, with one (or more) expert per system. Given the advanced technologies covered, the Summer School was aimed at PhD students or young post-docs who already had practical experience in confocal microscopy.

Topics included:

1) Lifetime analysis (FLIM) and super-resolved STED microscopy on a LEICA stellaris 8, excitation by white laser, with a STED section consisting of a depletion laser at 775nm, Tau STED and Gated STED modes and 3D STED modality. Tau and Gated STED can be used with the Tau Sense tool or with FALCON. This super-resolution modality enables fast acquisitions, with adapted fluorophores and relatively high phototoxicity.

2) Lifetime analysis (sFLIM) and dSTORM high-resolution microscopy on a Nikon eclipse TI/abbelight SAFE 360 combined with a spectral lifetime module (16 simultaneous channels) with a two-photon laser excitation tunable from 680 nm to 1080 nm. This super-resolution modality provides better spatial resolution, with longer acquisition times and up-to-date FLIM detection. 

3) Traditional confocal microscopy analysis combined with deconvolution on LEICA SP8 as a confocal microscopy control.

4) Use of Ratiometric biosensors  on LEICA SP5, fluorescent molecules that provide information about their environment (pH, ion concentration, ROS, etc.). Imaging them using a ratiometric approach is very widespread and has the great advantage of being suitable for all imaging systems (widefield, scanning confocal, spinning disk or super-resolution systems). In this practical session we looked at the prerequisites for this approach and the crucial points for accurate quantification (imaging parameters and image analysis methods).

5) STED/FLIM data analysis, on supplied datasets

6) STORM data analysis on supplied datasets

At the end of the Summer School, the experiences of all participants was confronted in a general discussion, to expose the differences and possible advantages of the imaging technologies approached.

> Participant flash-talks and poster session (5 hours): each participant gave a short “get to know me” flash talk at the beginning of the Summer School. The poster session was organized to allow each participant to discuss his/her poster and to see all posters.

> Social activities
A pétanque tournament (https://www.wikihow.com/Play-Petanque)
Dinner at a restaurant

This intensive and varied one-week program allowed many opportunities to discuss with speakers and fellow participants.

Organizers

Bertrand Dubreucq¹ (Coordinator)
Kalina Haas¹ (Coordinator)
Katia Belcram¹
Herman Höfte¹
Aurélie Hurel¹
Romain Le Bars²
Alexis Peaucelle¹
Alice Vayssières¹
Samantha Vernhettes¹

¹ Jean-Pierre Bourgin Institute - Plant Sciences / IJPB (Versailles, France)
² Institute of Integrative Biology of the Cell / I2BC (Gif-sur-Yvette, France)

Teaching language

English

Venue

Institut Jean-Pierre Bourgin - Plant Sciences
INRAE Centre Île-de-France Versailles-Saclay
Route de St-Cyr (RD 10)
78026 Versailles Cedex France

Sponsors