Multifocal multiphoton microscopy: a fast and efficient tool for 3-D fluorescence imaging

Multifocal multiphoton microscopy (MMM) is an efficient and technically sim ple method for generating multiphoton fluorescence images. Featuring the hi gh axial resolution of confocal and multiphoton scanning microscopes, MMM a lso achieves high speed in 3-D microscopy. In this paper, examples of fast- mode 3-D microscopy are given including imaging of fixed brain tissue as we ll as living PC12 cells. The imaging speed of MMM is solely determined by t he fluorescence photon Bur, so that in practice, for brightly fluorescent s pecimens, a whole stack of about 50 images of 30-70 mu m diameter can be ac quired within a few seconds. MMM represents a significant advance towards h igh speed nonlinear optical tomography of fluorescent specimens.