Endogenous SHG and 2PEF coherence imaging of substructures in neurons in 3D

Neuronal morphology, long-distance transport and signalling critically depend on the organization of microtubules in the cytoskeleton. Second harmonic generation (SHG) imaging has been recognized as a potentially powerful tool for in situ label-free neuroimaging with specific sensitivity to microtubules. We study here the structural organization of microtubules in living neurons using a wide-field multiphoton microscope that performs 3D imaging using a structured illumination. This microscope allows label-free high throughput imaging of living mammalian neurons. We show that we can image structural correlations by taking advantage of the structured illumination and the coherence of the emitted light. The result allows us to study the microtubule organization throughout the development of the neuron and to differentiate between the regions of the cytoskeleton in the matured neuron.