Two-Photon Neurotransmitter Uncaging for the Study of Dendritic Integration

  • Alexandra Tran-Van-Minh
  • Nelson Rebola
  • Andreas Hoehne
  • David A. DiGregorioEmail author
Part of the Neuromethods book series (NM, volume 148)


Neurons transform the information arising from up to thousands of synaptic inputs into specific spiking patterns. Nonlinear dendritic integration is a crucial step in this process and is thought to increase the computational ability of neurons. However, studying how complex spatiotemporal patterns of synaptic inputs drive neuronal spiking is technically challenging. Two-photon neurotransmitter uncaging allows researchers to activate sequences of single synapses with high spatiotemporal precision and thus systematically examine how single and multiple synaptic activation patterns may recruit dendritic nonlinearities. Here, we describe the theoretical and practical considerations of using two-photon uncaging to mimic synaptic activation and monitor the electrical and biochemical signaling in dendrites when evoked by various synaptic patterns.


Two-photon microscopy Neurotransmitter uncaging Dendritic integration Synaptic potentials Photolysis Neuronal computations 



Glutamate concentration


Two-photon laser scanning microscopy


Artificial cerebrospinal fluid


α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor


Acousto-optic deflector


Acousto-optic modulator




Dendritic input/output relationship


Two-component optical voltage sensor made of the neuronal tracer dye DiOC16(3) and dipicrylamine

DNI-glutamate TFA

4-Methoxy-5,7-dinitroindolinyl-L-glutamate trifluoroacetate


The molecular two-photon cross-section is usually quoted in the units of Göppert-Mayer (GM), where 1 GM is 10−50 cm4 s photon−1


Input/output relationship




N-Methyl-D-aspartate receptor


Point spread function




Spatial light modulator


Uncaging-evoked glutamate transient


Uncaging-evoked EPSC


Uncaging-evoked EPSP


Voltage-gated calcium channel


Bragg angle, the angle between the incident laser beam into an AOM and the diffracted beam


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Alexandra Tran-Van-Minh
    • 1
    • 2
  • Nelson Rebola
    • 1
    • 2
  • Andreas Hoehne
    • 1
    • 2
  • David A. DiGregorio
    • 1
    • 2
    Email author
  1. 1.Laboratory of Dynamic Neuronal Imaging, Department of Neuroscience, Institut PasteurParisFrance
  2. 2.CNRS UMR 3571ParisFrance

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