Multi-Photon-Laser-Scanning-Microscopy (MPLSM) now stands as one of the most powerful experimental tools in biology. Specifically, MPLSM based in-vivo studies of structures and processes in the brains of small rodents and imaging in brain-slices have led to considerable progress in the field of neuroscience. Equipment allowing for independent control of two laser-beams, one for imaging and one for photochemical manipulation, strongly enhances any MPLSM platform. Some industrial MPLSM producers have introduced double scanner options in MPLSM systems. Here, we describe the upgrade of a single scanner MPLSM system with equipment that is suitable for independently controlling the beams of two Titanium Sapphire lasers. The upgrade is compatible with any actual MPLSM system and can be combined with any commercial or self assembled system. Making use of the pixel-clock, frame-active and line-active signals provided by the scanner-electronics of the MPLSM, the user can, by means of an external unit, select individual pixels or rectangular ROIs within the field of view of an overview-scan to be exposed, or not exposed, to the beam(s) of one or two lasers during subsequent scans. The switching processes of the laser-beams during the subsequent scans are performed by means of Electro-Optical-Modulators (EOMs). While this system does not provide the flexibility of two-scanner modules, it strongly enhances the experimental possibilities of one-scanner systems provided a second laser and two independent EOMs are available. Even multi-scanner-systems can profit from this development, which can be used to independently control any number of laser beams.

Date Published: 26 February 2010
PDF: 7 pages
Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 75692Y (26 February 2010); doi: 10.1117/12.842156

Published in SPIE Proceedings Vol. 7569:
Multiphoton Microscopy in the Biomedical Sciences X
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)