Plateforme Intégrée de Biophysique et de Biologie Structurale



Managers Emmanuel Margeat , Antoine Le Gall

Measure conformational changes, interactions, changes in the environment of single molecules in milliseconds to seconds timescale. Measure fast (milliseconds timescale) two and three dimensional localization of molecules of interest.

TIRF with dual-view on one side => Single particle tracking, single molecule FRET with Alex ( Alternating-laser excitation)
3D-MFM on the other side => Multi-focal microscopy (3D imaging and tracking)

Specifications TIRF/HILO/Epifluorescence wide field configuration. Dual color smFRET/SPT and 3D imaging and tracking
  • Inverted microscope (Zeiss Axiovert 200)
  • Zeiss alpha Plan-Apochromat, 100X, NA1.46
  • Band-pass filters to select excitation wavelengths (450, 488, 514, 532, 640 nm)
  • Laser diodes 450, 488, 525 nm, Thorlabs PL450B, L488P60 and PL520
  • Laser 532nm,
  • Laser 640 nm, spectra physics, EXLR-640C-100-CDRH
All laser lines are controlled through an AOTF (AOTFnC-400.650-TN AAoptics, France) and can be set to TIRF/HILO/Epifluorescence illumination mode
MFM setup:
  • First relay lense L1 (f = 150 mm(to form a secondary Fourier plane, conjugate of the objective pupil plane)
  • Custom-built multifocus grating (MFG)
  • Chromatic correction grating (CCG)
  • Multifacet prism, combined with a second relay lens L2 (f = 200mm)
  • emCCD camera (IXON DU-897, 512*512, 16µm, Andor, Ireland)
optional : cylindrical lens in front of the em-CCD (for 3D single molecule localization using astigmatism).