Multiphoton microscopy is a powerful tool in neuroscience, promising to deliver important data on the spatiotemporal activity within individual neurons as well as in networks of neurons. A major limitation of current technologies is the relatively slow scan rates along the z direction compared to the kHz rates obtainable in the x and y directions. Here, we describe a custom-built microscope system based on an architecture that allows kHz scan rates over hundreds of microns in all three dimensions without introducing aberration. We further demonstrate how this high-speed 3D multiphoton imaging system can be used to study neuronal activity at millisecond resolution at the subcellular as well as the population level.
Proc Natl Acad Sci U S A
2919 - 2924
Animals, Cerebral Cortex, Imaging, Three-Dimensional, Mice, Mice, Inbred C57BL, Microscopy, Fluorescence, Multiphoton, Neurons, Reproducibility of Results