At the Caltech Biological Imaging Center

Main components of the TPLSM

The microscope is a modified Molecular Dynamics Sarastro 2000 confocal, with a Nikon Optiphot 2 upright microscope. Only minor modifications were made to allow 2-photon imaging, and the ability to do standard confocal imaging has been retained. 2-Photon imaging is carried out using the Coherent Mira 900 mode-locked Ti:sapphire IR laser, pumped by a Coherent Innova 310 8W argon-ion laser.

The two periscope mirrors that bring the IR beam up to the optical table of the Sarastro 2000, as well as the two mirrors that bounce the IR beam from the Mira 900 to the periscope (see Light Path), are optimized to reflect IR. All of the laser beams are enclosed in metal tubes and boxes to allow safe operation even by biologists.

Not shown are the pump laser power supply and heat exchanger, Mira 900 controller, scanner controller box, and Silicon Graphics Indigo computer.

The Molecular Dynamics ImageSpace software controls the Sarastro 2000, and does all the data acquisition, image processing, manipulation and quantification of data.