Four-dimensional multi-site photolysis of caged neurotransmitters

Mary Go, Minh-Son To, Christian Stricker, Stephen Redman, Hans-A Bachor, Greg Stuart, Vincent Daria

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    25 Citations (Scopus)


    Neurons receive thousands of synaptic inputs that are distributed in space and time. The systematic study of how neurons process these inputs requires a technique to stimulate multiple yet highly targeted points of interest along the neuron's dendritic tree. Three-dimensional multi-focal patterns produced via holographic projection combined with two-photon photolysis of caged compounds can provide for highly localized release of neurotransmitters within each diffraction-limited focus, and in this way emulate simultaneous synaptic inputs to the neuron. However, this technique so far cannot achieve time-dependent stimulation patterns due to fundamental limitations of the hologram-encoding device and other factors that affect the consistency of controlled synaptic stimulation. Here, we report an advanced technique that enables the design and application of arbitrary spatio-temporal photostimulation patterns that resemble physiological synaptic inputs. By combining holographic projection with a programmable high-speed light-switching array, we have overcome temporal limitations with holographic projection, allowing us to mimic distributed activation of synaptic inputs leading to action potential generation. Our experiments uniquely demonstrate multi-site two-photon glutamate uncaging in three dimensions with submillisecond temporal resolution. Implementing this approach opens up new prospects for studying neuronal synaptic integration in four dimensions.

    Original languageEnglish
    Number of pages9
    JournalFrontiers in Cellular Neuroscience
    Issue numberDEC
    Publication statusPublished - 2 Dec 2013


    • Caged neurotransmitters
    • Holographic projection
    • Synaptic integration
    • Two-photon microscopy
    • Two-photon photolysis


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