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Cellular imagery

The d'Alembert Institute imaging platform gives access to various biological and biophysical, intra and intercellular studies for all kind of cells (plant, mammalian, bacterial). The different microscopies are adapted to the multi-disciplinary research developed at the d'Alembert Institute.

The cellular imaging platform is spread over two rooms. A confocal microscope Leica SP2 is housed in a cellular culture room (L2). A Leica SP8 and a Zeiss AxioObserver microscope with a TIRF module is housed in the d'Alembert Institute.

  • The confocal Leica SP2 is equipped with a temperature and CO2 controller for the observation of living cells. Different excitation wavelengths are available (458, 476, 488, 514, 543 et 633 nm). FLIM (Fluorescence Lifetime Imaging Microscopy) with a Becker Hickel 830 card are possible. A pulsed femtosecond Mai-Tai allows biphotonic excitation between 720 and 920 nm. Several detectors are available: three analogic and one single photon detectors. Also, three objectives 10x,63x,100x are present. FLIM measurements are possible to study biomolecule interactions in cellulo by FRET (Förster resonance energy transfer). The femtosecond laser allows photoactivation by multiphoton excitation (IR) of biological processes. For instance, we are studying the photo-controlled triggering of electron transfer occuring at the NO Synthase level targeted by photoactivable NADPH analogues. We are also investigating the apoptotic processes triggered by two-photon absorption compounds targeting mitochondria.
  • The confocal SP8 is equipped with two analogic and one hybrid detectors which allows quantitative measurements. The available excitation wavelengths are 405, 458, 476, 488, 514, 543 et 633 nm. Three objectives are present (10x, 63x and 100x). Phase contrast images can also be obtained.
  • The Zeiss AxioObserver TIRF microscope gives the opportunity to study dynamic events at high spatial and temporal resolution at the proximity of the coverslip (about 200 nm). A EMCCD Evolve 512 gives access to very fast imaging (>100 frames/sec). Two Laser diodes 450 and 560 nm area available to excite the most used fluorophores.

Numerous and very diverse themes are investigated: cancer (Biphenol A effect on cancerous cells), pharmacology, bioenvironment (bacterial biofilm dispersion), new biofuel generation (lipid synthesis by microalgae), traffic and localisation intracellular (small GTPases), photodynamic therapy (cell death triggered by multiphotonic activation).

Realisation examples:

  • Chennoufi R. et al. "Mitochondria-targeted Triphenylamine Derivatives Activatable by Two-Photon Excitation for Triggering and Imaging Cell Apoptosis." Sci. Rep. (2016) 6:21458.
  • Chennoufi R. et al. "Differential Behaviour of Cationic Triphenylamine Derivatives In Fixed and Living Cells." Chem. Commun. (2015) 51, 14881-14884.
  • Li Y. et al. "Rational design of a fluorescent NADPH derivative imaging constitutive nitric-oxide synthases upon two-photon excitation." Proc. Natl. Acad. Sci. U.S.A (2012) 109, 12526-31.

Equipments list :

-  Confocal Leica SP2 + laser femtoseconde
- Confocal Leica SP8
- Zeiss AxioObserver avec module TIRF.

Localisation :

- d'Alembert building 2nd floor, room  A2-04

- IDA building , ground floor, room  H9