In vivo two-photon microscope with a highly flexible frontend for in vivo research.

The custom design ensures fast and reproducible access to the area of interest, including rotation of imaging plane, and maximizes space for auxiliary experimental equipment in the vicinity of the animal. Mechanical flexibility is achieved with large motorized linear stages that move the objective in the X, Y, and Z directions up to 130 mm. +- 45° rotation of the frontend (rotational freedom for one axis) is achieved with the combination of a motorized high precision bearing and gearing. The high-speed two-photon microscope is able to scan large volumes at high speeds. Resonant scanning technology is used to produce two-dimensional (x,y) images with high frame rates (512x512 pixels at 30 Hz). A tuneable lens system is used to quickly shift the focus in z dimension. The combination of these two systems is able to scan 3D volumes in-vivo (1 x 1 x 0.5mm at 8 Hz).

Applications:

In-vivo time-lapse two-photon imaging
Photoactivation/ -conversion
Volume scanning
Imaging of weak fluorescence signals

Objectives
magnif.	NA	type	specific
16 x	0.8	NA 3 mm	Water immersion

Lasers

Tunable Ti:Sa femtosecond laser

(80 Mhz, 140 fs, 680-1060 nm)(Chameleon Ultra II, Coherent)

Femtosecond fiber laser (80 Mhz, 150 fs, 1064 nm)(FemtoPower FP-1060-5-fs, Fianium)

detectors.

filter or beamsplitter

2 GaAsP-PMTs

GFP 500 - 540

FP BP 565 - 635

Software:

Scanimage 5.2

Computer:

Core i5 16GB RAM