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A unique tool for characterizing interactions |
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+33 5 82 52 58 07Streak-FLIM (Fluorescence Lifetime Imaging Microscopy) is a unique technique developed by the FRAIB imaging platform, which makes it possible to reveal protein-protein and protein-DNA interactions, or changes in spatial conformation. If they are close enough to interact, molecules transfer energy to each other and this can be measured through fluorescence lifetime, providing a wealth of useful information. These measures are nevertheless independent of the concentration of the molecule and its fluorescence intensity.
Services:
- Detection of molecular interactions (protein – protein, protein – DNA, etc.) by measuring the fluorescence lifetime of the donor in the absence or presence of an acceptor.
- Detection by a Streak camera (Hamamatsu Streakscope) for measuring the lifetime, with a resolution on the order of a picosecond.
- Variable time acquisition (from seconds to tens of seconds) according to the fluorescence intensity and the type of information sought (lifetime by ROI, or pixel-based).
Access mode:
- With assistance
- As collaborative project
- As service delivery
Technical characteristics:
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- Nikon TE2000 inverted microscope
- Images in x, t (t in variable full scale according to the voltage ramp applied to the deflection electrodes from 1 ns to 1 µs).
- Images in x, t with stacks of images in y.
- Obtaining the fluorescence decay curves for one ROI or per pixel.
- Determining the ai and ti parameters by least squares adjustment of the obtained experimental curves (nonlinear least squares or maximum likelihood).
Available resources
Name |
Contact |
Location |
Two-photon FLIM
One-photon FLIM
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Cécile Pouzet
+33 5 82 52 58 07 |
FRAIB – Campus INRA Auzeville |
Software manual
- Download the “Topography” software manual dedicated to streak-FILM
- “Taupography” is made by FRAIB imagery platform and LRSV lab. This software is under the licence of Photonics, so please contact Thomas Gelot or Eric Dréan.
Some publications made thanks to these resources:
- A newly evolved chimeric lysin motif receptor-like kinase in Medicago truncatula spp. tricycla R108 extends its Rhizobia symbiotic partnership [Academic Journal] – Luu, Thi-Bich; Ourth, Anna; Pouzet, Cécile; Pauly, Nicolas; Cullimore, Julie – New Phytologist. Sept. 2022, Vol. 235 Issue 5, p1995, 2007 p. DOI: 10.1111/nph.18270
- Ormancey, M., Guillotin, B., Merret, R. et al. Complementary peptides represent a credible alternative to agrochemicals by activating translation of targeted proteins. Nat Commun 14, 254 (2023). >>>
- Camborde L, Kiselev A, Pel MJC, Le Ru A, Jauneau A, Pouzet C, Dumas B, Gaulin E. An oomycete effector targets a plant RNA helicase involved in root development and defense. New Phytol. 2022 Mar;233(5):2232-2248. doi: 10.1111/nph.17918. Epub 2022 Jan 7. PMID: 34913494.
- Diana Ramirez-Garcés, Laurent Camborde, Michiel Pel, Alain Jauneau, Yves Martinez, et al.. CRN 13 candidate effectors from plant and animal eukaryotic pathogens are DNA ‐binding proteins which trigger host DNA damage response. New Phytologist, 2016, 210 (2), pp.602-617. ⟨10.1111/nph.13774⟩. ⟨hal-03807803⟩
- Laurent Camborde, A. Jauneau, Christian Brière, Laurent Deslandes, Bernard Dumas, et al.. Detection of nucleic acid-protein interactions in plant leaves using fluorescence lifetime imaging microscopy. Nature Protocols, 2017, 12 (9), pp.1933-1950. ⟨10.1038/nprot.2017.076⟩. ⟨hal-01607375⟩
- Dorian Farache, Alain Jauneau, Cécile Chemin, Marine Chartrain, Marie-Hélène Rémy, Andreas Merdes and Laurence Haren. Functional analysis of Gamma-tubulin Complex Proteins indicates specific lateral association via their N-terminal domains. JBC, 2016
- A. Testard, D. Da Silva, M Ormancey, C. Pichereaux, C. Pouzet, A. Jauneau, S. Grat, E. Robe, Brière, V. Cotelle, C. Mazars and P. Thulleau. Calcium- and Nitric Oxide-Dependent Nuclear Accumulation of Cytosolic Glyceraldehyde-3-Phosphate Dehydrogenase in Response to Long Chain Bases in Tobacco BY-2 Cell. Plant Cell Physiol, 2016
- Marçal Soler, Anna Plasencia, Romain Larbat, Cécile Pouzet, Alain Jauneau, Susana Rivas, Edouard Pesquet, Catherine Lapierre, Isabelle Truchet and Jacqueline Grima-Pettenati. The Eucalyptus linker histone variant EgH1.3 cooperates with the transcription factor EgMYB1 to control lignin biosynthesis during wood formation. New Phytologist, 2016
- Judith Fliegmann, Alain Jauneau, Carole Pichereaux, Charles Rosenberg, Virginie Gasciolli, Antonius C.J. Timmers, Odile Burlet-Schiltz, Julie Cullimore and Jean-Jacques Bono. LYR3, a high-affinity LCO-binding protein of Medicago truncatula, interacts with LYK3, a key symbiotic receptor. Febs Letters, 2016
- Clementine Le Roux,Gälle Huet, Alain Jauneau, Laurent Camborde, Dominique Tremousaygue, Alexandra Kraut, Binbin Zhou, Marie Levaillant, Hiroaki Adachi, Hirofumi Yoshioka, Sylvain Raffaele, Richard Berthome, Yohann Coute, Jane E. Parker,and Laurent Deslandes. A Receptor Pair with an Integrated Decoy Converts Pathogen Disabling of Transcription Factors to Immunity. Cell, 2015
- Josep Vilarrasa-Blasi, Mary-Paz Gonzalez-Garcıa, David Frigola, Norma Fabregas-Vallve, Konstantinos G. Alexiou, Nuria Lopez-Bigas, Susana Rivas, Alain Jauneau, Jan U. Lohmann, Philip N. Benfey, Marta Ibanes, and Ana I. Cano-Delgado Regulation of Plant Stem Cell Quiescence by a Brassinosteroid Signaling Module. Developmental Cell, 2014
- Stella Cesari Gaëtan Thilliez, Cécile Ribot, Véronique Chalvon, Corinne Michel, Alain Jauneau, Susana Rivas, Ludovic Alaux, Hiroyuki Kanzaki, Yudai Okuyama, Jean-Benoit Morel, Elisabeth Fournier, Didier Tharreau, Ryohei Terauchi and Thomas Kroj. The Rice Resistance Protein Pair RGA4/RGA5 Recognizes the Magnaporthe oryzae Effectors AVR-Pia and AVR1-CO39 by Direct Binding. The Plant Cell, 2013, 25:1463-1481
- Daniel Marino, Solène Froidure, Joanne Canonne, Sara Ben Khaled, Mehdi Khafif, Cécile Pouzet, Alain Jauneau, Dominique Roby, Susana Rivas. Arabidopsis ubiquitin ligase MIEL1 mediates degradation of the transcription factor MYB30 weakening plant defence. Nature Communications, 2013, 4: 1476 .