2-Photon Imaging Resources

The 2-photon core is based on two imaging systems from Wisconsin-based Prairie Technologies, Inc. Each has a separate workstation, and depending on laser settings the two systems can often be used simultaneously by independent users.

One workstation is built around a modified Zeiss Axio Examiner z1 microscope that is configured for brain slice imaging and physiology (left). The second workstation is based on a custom-modified Olympous BX-61WIF microscope suited for in vivo imaging of living brain (below left). Both microscopes use Zeiss objectives, including the incomparable 20x 1.0NA DIC (UV) VIS-R objective. Each objective can be used in conjunction with a piezo Z-axis drive for high-speed and high-precision Z-movements.

The two workstations share two separate Mai Tai DeepSee Ti:sapphire lasers, both of which are tunable from 690 -1020 nm and are configured for two-photon imaging or uncaging. There are also two additional lasers, one at 471 nm and the other at 563 nm, suitable for single-photon uncaging. The beams can be split such that any of the four lasers can be directed to either workstation.

The power of this unique imaging system is that the two Mai Tai lasers can be used simultaneously, allowing dual two-photon imaging at two wavelengths, or two-photon imaging at one wavelength combined with two-photon uncaging at a different wavelength. Either setup can be further combined with simultaneous single-photon uncaging at either 471 or 563 nm (or both in combination).

Scanning can be driven either by a galvanometer or (for high-scan speeds) by an acousto-optical deflector (AOD).

The core also has five licenses for Neurolucida software, including the Autoneuron extension module. Neurolucida permits precise 3D tracing, mapping and reconstruction of neurons and their morphology from microscopic images. Autoneuron supports automated or semi-automated reconstruction of neuronal processes.