The technology of multiphoton microscopy allows imaging of visible light fluorophores within a living animal from the brain surface to a depth approximating layer III of mouse cortex.
Scanning occurs in X, Y and Z directions for 3-dimensional rendering. Viral delivered fluorescent markers and small molecule fluorescent reporters are routinely used. The unit consists of a BioRad1024 multiphoton microscope with custom modifications mounted on an Olympus BX50WI upright microscope.
A mode-locked Ti:Saphire laser (MaiTai) generates two-photon fluorescence with 800 nm excitation, and detectors containing three photomultiplier tubes (Hamamatsu Photonics) collect emitted light in the ranges of 380-480, 500-540, and 560-650 nm. The microscope stage has been modified to allow stereotactic frame placement, inhalant anesthesia, and temperature control.
A surgical suite is located adjacent to the microscope facility and is equipped with all necessary tools for implantation of cranial windows for imaging.