LSM 710 confocal microscope + Two photon system
The Zeiss LSM 710 laser scanning confocal microscope features a 32-channel linear detector for spectral analysis and two flanking photomultiplier (PMT) channels that can be combined to monitor dynamic changes of multiple fluorophores. Modules for advanced confocal applications such as Fluorescence Energy Transfer (FRET), colocalization analysis and Recovery After Photobleaching (FRAP) are available. The multi-time plug in allows automated 3D (x,y,z) or 4D (x,y,z,t) scans of preprogrammed locations and 3D image stitching. The laser lines include: 405 nm diode laser (Dapi), 458 nm, 488 nm and 514 nm multiline Argon laser, 561 nm diode pumped solid state laser and 633 nm HeNe laser. The objectives are: 10x dry, 25x W/oil/glycerin, 40x dry/oil and 63x oil.
Two-photon microscopy is based on the principle of two-photon excitation by a pulsed near-infrared (NIR) laser and enables fluorescence imaging of optical sections with subcellular resolution. NIR light penetrates deeper into scattering tissue, allowing imaging deeper in intact samples. Image stacks of defined depth ranges can be used for spatial reconstruction to visualize fluorescent structures in 3D. Two-photon microscopy is more advantageous then other microscopic techniques for visualizing structures deeper in scattering samples (tissues) in three dimensions and reduced photobleaching outside of the focal plane.
