Use of Multiphoton Imaging for Studying Cell Migration in the Mouse

We describe a method for studying cell motility in the living mouse using multiphoton microscopy. The procedure consists of mouse anesthesia, labeling of target cells with enhanced green fluorescent protein by infection with recombinant adenovirus, implantation of beads carrying chemoattractant, preparation of the mouse for imaging, and imaging of individual cell motions via multiphoton microscopy. Two-photon fluorescence excitation of enhanced green fluorescent protein allows visualization of cells within the dermis, whereas second harmonic generation (a non-linear scattering process) allows a simultaneous detailed definition of the dermis structure.