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Institut Pasteur Course: Principles and Applications of Fluorescence Microscopy 2017-2018


PRINCIPLESANDAPPLICATIONSOFFLUORESCENCEMICROSCOPY

26 February – 9 March 2018

DIRECTORS

David DIGREGORIO Jost ENNINGA Gaël MONERON Institut Pasteur Institut Pasteur Institut Pasteur

HEAD OF LABORATORY

Florian RÜCKERL Institut Pasteur

Teaching Assistants

Laurent AUDRY Institut Pasteur

Mickael LELEK Institut Pasteur

Patricia LATOUR-LAMBERT

Institut Pasteur

Joana LOURENCO

ICM - INSERM

Nelson REBOLA ICM

Cécile SAINT CLOMENT

Institut Pasteur

Jean-Yves TINEVEZ Institut Pasteur

Julie NGUYEN CPN-INSERM

Diana ZALA CPN- INSERM

Marie-Pierre ADAM Institut Pasteur

Min Hee KIM Institut Pasteur

Olivier DISSON Institut Pasteur

Course Staff

Guylène K’OUAS Institut Pasteur

Gauthier PUIME Institut Pasteur

Hervé WAXIN Institut Pasteur

Location Education Center – Institut Pasteur – Pavillon Louis Martin

28, rue du Docteur Roux 75015 Paris Lectures: Room No 2; Practical Work: Room on the ground floor

With the participation of:


WEEK1 Monday, 26 February 8:00 – 8:30 Badges 8:30 – 9:30 Welcome, Admin, and Introduction David DIGREGORIO

Jost ENNINGA Gaël MONERON

(Institut Pasteur, France) 9:30 – 12:00 Basic optics I Gaël MONERON

• Geometrical and Gaussian optics (Institut Pasteur, France) • Basic principles of a light microscope • Contrast techniques

13:30 – 18:30 Bench optics practicals 19:00 Welcome drink Tuesday, 27 February 9:00 – 12:00 Basic optics II Gaël MONERON

• Coherence of light (Institut Pasteur, France) • Diffraction, Fourier optics, Resolution • Objective lenses and aberrations

13:30 – 18:30 Bench optics practicals Wednesday, 28 February 9:00 – 12:00 Fluorescence, light sources Gaël MONERON

• Photo-physics (Institut Pasteur, France) • Multi-color imaging strategies • Lamps, LEDs, LASERs, wavelength selection devices

13:30 – 14:30 Imaging modalities, detectors David DIGREGORIO

• Wide field / beam Scanning (Institut Pasteur, France) • Cameras / point detectors • Signal-to-noise ratio (sources of noise) • Sampling (time and space)

14:30 – 18:30 Bench optics practicals


Thursday, 1 March 9:00 – 9:45 Confocal laser scanning microscopy David DIGREGORIO

(Institut Pasteur, France) 9:45 – 10:30 Dynamic imaging David DIGREGORIO

(Institut Pasteur, France) 10:30 – 12:00 Multi-photon microscopy Florent HAISS

(Institut Pasteur, France) 13:30 – 14:00 Estimating resolution (PSF) David DIGREGORIO (Institut Pasteur, France) 14:00 – 17:00 PSFs practicals on commercial microscopes 17:00 – 18:30 Wrap-up Friday, 2 March 9:00 – 10:30 Biosensors Jost ENNINGA

(Institut Pasteur, France) 10:30 – 12:00 New fluorescent probes Marcel BRUCHEZ (Carnegie Mellon Univ., USA) 13:30 – 15:00 Structured illumination microscopy Alexandra FRAGOLA for enhanced resolution imaging (Univ. Pierre & Marie Curie, France) in biology 15:00 – 15:30 Imaging infection Jost ENNINGA (Institut Pasteur, France) 15:30 – 18:30 Fixed preparation imaging practicals

• Fluorescent neurons • Microtubules • Bacterial invasion

Saturday, 3 March 9:00 – 12:00 Introduction to image analysis Jean-Yves TINEVEZ for Life-Science microscopy (Institut Pasteur, France) 13:00 – 18:00 Image analysis practicals

• Image analysis • Data presentations


WEEK2 Monday, 5 March 9:00 – 10:30 Single molecule localization Christophe ZIMMER microscopy principles (Institut Pasteur, France) 10:30 –12:00 TIRF and SAF microscopy Sandrine LEVEQUE-FORT

(Univ. Paris-Sud, France) 13:30 – 15:00 STED Microscopy Gaël MONERON (Institut Pasteur, France)

15:00 – 18:30 Fixed preparation imaging practicals • Fluorescent neurons • Microtubules • Bacterial invasion • Organelles

Tuesday, 6 March 9:00 – 10:30 FRAP/Photoactivation Jeremy DITTMAN

(Weill Cornell Medical College, USA) 10:30 – 12:00 FCS/FLIM Christian EGGELING

(Oxford University, UK) 13:30 – 15:00 Single molecule/FRET/TIRF Keith WENINGER

(North Carolina State, USA) 15:00 – 18:30 Live cell imaging practicals

• XFP neurons (culture, acute brain slices) • Live Bacteria invasion • Organelles

Wednesday, 7 March 9:00 – 10:30 High Content Screening Nathalie AULNER (Institut Pasteur, France) 10:30 – 12:00 Imaging the immune system Philippe BOUSSO (Institut Pasteur, France) 13:30 – 15:00 Light-sheet fluorescence microscopy Willy SUPATTO

(Ecole Polytechnique, France) 15:00 – 18:30 Live cell imaging practicals

• Calcium imaging (culture, acute brain slices) • Rac Photoswitching • FRAP, paGFP photoactivation


Thursday, 8 March 9:00 – 10:30 Imaging deep in scattering tissue Sylvain GIGAN (Univ. Paris Pierre et Marie Curie, France) 10:30 – 12:00 Wrap-up and Analysis 13:30 – 18:30 Live cell imaging practicals

• Calcium imaging (culture, acute brain slices) • Rac Photoswitching • FRAP, paGFP photoactivation

Friday, 9 March 9:00 – 17:30 Analysis and final presentations 17:30 – 18:30 Written exam and course evaluation 19:00 Aperitif/Dinner/Social


Administration Institut Pasteur Education Center - 28, rue du docteur Roux Building 9C - Pavillon Louis Martin Isabelle DI MARTINO Tél : +33 (0)1 45 68 82 89 EMAIL : [email protected] [email protected] Registrar’s Office 28, rue du docteur Roux Building 13 - Salle Saint-Joseph de Cluny Sylvie MALOT Tél : +33 (0) 45 68 81 41 EMAIL : [email protected]

institut pasteur course: principles and applications of ... · for life-science microscopy...

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