Contact : Marie Erard
BIFLUOR
Multimodal fluorescence spectromicroscopy
The BIFLUOR microscope is a home-made multimodal set-up combining different modes of fluorescence analyses inside living cells (see below). It is charaterized by a very high sensitivity FLIM system based on non-descanned (non-confocal) fluorescence detection.
• Fluorescence Lifetime Imaging Microscopy (FLIM)
Excitation : pulsed laser diodes (442nm et 466nm, 80ps, 20MHz)
Fiber-coupled laser scanning head C1 Nikon
Fast MCP-PMT detector (Hamamatsu) and TCSPC analysis (PicoQuant)
• Quantitative fluorescence intensity imaging
Excitation : HBO lamp
Semrock & Chroma filter cubes (CFP, YFP, GFP, TRITC, Cherry, DAPI)
Orca-AG CCD camera
• Spectral analysis
Fiber-coupled Ocean Optics JAZ system 250nm-800nm
Objectives :
Nikon x63 1.2 NA water immersion FI plan-apochromatic
Nikon x40
Sample : Attofluor cell chamber, coverslips diam 25mm
Temperature control : 20°-40°C
Applications :
• membrane biophysics (1)
• pH & ROS sensing (2)
• FRET biosensors (3)
• protein-protein interactions (4)
• intracell. luminescence of gold NPs (5)
(1) Boucherie et al 2013 Biol of the Cell 105 561
(2) Poea Guyon et al 2013 Anal. Bioanal. Chem. 405 3983
Poea Guyon et al 2013 J. Cell Biol. 203 283
Dupré-Crochet et al 2013, J. Leukoc. Biol. 94 657
(3) Erard et al 2013 Mol Biosystems 8 258
Betolngar et al 2015 Anal Bioanal Chem 407 4183
(4) Wade et al 2011 J Biol Chem 286 15252
C. Ziegler Thèse IDEX Paris-Saclay 2012-2016
(5) Yahia-Ammar et al (2016) ACS Nano in press