22 Oct 2-photon imaging on the Bordeaux Imaging Center
Two systems are available on the Photonic Unit of the Bordeaux Imaging Center to perform 2-photon microscopy:
1) our SP5-MP Confocal microscope is equipped, in addition to conventional visible lasers, with a femtosecond laser Mai Tai HP (Spectra Physics) to excite fluorochromes from 690 to 1040 nm. The detection is done by 2 NDD hybrid detectors to collect the maximum of fluorescence signal.
Moreover, a specific filters set and a transmission detector allows to acquire Second Harmonic Generation (SHG) signal to observe high organized and non-centrosymmetric structures like collagen or myosin without any staining.
A Tokai Hit stage insert can be installed to keep your sample alive by maintaining the temperature and the concentration in CO2.
The possibility to switch between 2 scanners (galvanometric or resonant) brings a very high flexibility for the different kind of experiments.
Finally, a TCSPC FLIM module from PicoQuant driven by the software SymphoTime64 allows to perform lifetime experiments to measure FRET for instance.
2) Our Femtonics system is dedicated to imaging on thick tissue slices or entire animal (mouse and rat but also drosophila or zebrafish …).
To do that, two femtosecond lasers are available:
– A Chameleon Vision 2 (Cohérent) is used for imaging, couple with galvo or resonant scan head depending on the experiment. Its wavelength range, from 690 to 1080 nm with precompensation, allows very efficient excitation of green fluorescent fluorochromes like GFP but also GCaMP6f (calcium sensor) or iGluSnFR (glutamate sensor), and some red fluorophores (dTomato).
– A Mira 900 is dedicated to photo-manipulation through the galvo scan head. This second pathway offers a very high flexibility to our system to perform uncaging or optogenetic simultaneously with imaging.
The detection is done by 5 GaAsP detectors (2 upper and 2 lower, 1 for transmitted light) to collect a maximum amount of light.
If you want to acquire images of very thick samples, fixed or alive, with or without staining, don’t hesitate to contact us. A test session on one for these microscopes is possible!
