First "real" use of Prairie Technologies' Ultima dual 2-photon microscope.

Early in 2001 Haruo Kasai and I were starting to write a NIH grant together on using 2-photon uncaging and he revealed that his lab made their first dual 2-photon microscope. This was months before we even submitted our Nature Neuroscience paper on the first diffraction-limited 2-photon uncaging of glutamate. I was rather shocked, and I remember asking if I had read that correctly: "Do you mean two two-photon lasers?!" Apparently my eyes were not decieving me, and I thought I have got to get one. So I approached Prairie Technologies (in which I have no financial interest, by the way) to make a new scan head with two sets of galvonometers and no pin holes, designed specifically to control two Ti:sapphire lasers independently. In order to cover the initial development cost, I had to find a second chump who was willing to take a chance on this venture into the unknown. That chump was Jeff Magee (then at the Neuroscience Center, LSU). Thus, Prairie made two dual 2-photon scan heads, which they dubbed "The Ultima". Jeff got his first (as he had 2 lasers in place) and mine came a little later in May 2003 (Prairie got official permission from BioRad to make these two instruments, as BioRad held the 2P patent at that time). Independently of all this, during 2002 Bernardo Sabatini had established his new lab Neurobiology at Harvard. He had built his own dual 2P laser single scan head microscope, based on the hardware and software made in the Svoboda Lab at CSH. Thus, the race was on to produce the first sexy experiments with dual 2P microscopes. It turned out that the Kasai lab won that race with our Nature paper in 2004. Sabatini was the first (just) to publish 2P Ca imaging with 2P uncaging of glutamate in his October 2004 paper in Neuron (parenthetically, I would like to point out that in his News & Views of this paper, Nelson Spruston seems to deliberately (mis)cite Jackie Schiller's UV uncaging paper from 1998 as "2-photon laser uncaging" (so forming the basis for Sabatini own application), rather than our 2001 Nature Neuroscience paper). We published our first Ca/glutamate paper in Neuron in May 2005.

In Fabruary and April 2006 the Magee lab published two papers using the Ultima to its full extent. In both papers they do rapid multi-site 2P uncaging of MNI-glutamate in elegant studies of dendritic summation (the J Neurosci paper uses 2P uncaging to reproduce (amazingly) the output patterns of rat CA1 neurons during (a) eye movement in sleeping and (b) environmental exploration). Mike Hausser wrote a nice News & Views for the Neuron paper (the picture is taken from that article). I want to thank Mike for citing our Nature Neuroscience paper and congratulate Jeff on some fantastic work. Good luck Jeff at Janelia Farm!

Gasparini S, Magee JC (2006) State-Dependent Dendritic Computation in Hippocampal CA1 Pyramidal Neurons. J. Neurosci. 26: 2068-2100.

Losonczy1 A, Magee JC (2006) Integrative Properties of Radial Oblique Dendrites in Hippocampal CA1 Pyramidal Neurons. Neuron 50: 291–307.

Matsuzaki M, Tachikawa A, Ellis-Davies GCR, Miyashita Y, Iino M, Kasai H (2001) Dendritic spine morphology is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurones. Nature Neurosci. 4: 1086-1092.

Matsuzaki M, Honkura N, Ellis-Davies GCR, Kasai H (2004) Structural basis of functional synaptic plasticity in single dendritic spines. Nature 429: 761-766.

Noguchi J, Matsuzaki M, Ellis-Davies, GCR, Kasai, H. (2005) Spine-neck geometry determines NMDA-receptor dependent Ca signaling in dendrites. Neuron 46, 609 -622.

Carter AG, Sabatini BL (2004) State-dependent calcium signaling in dendritic spines of striatal medium spiny neurons. Neuron 44: 483–493.

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