Members: Dr. Gergely KATONA, Dr. Gábor JUHÁSZ, Dr. Dénes PÁLFI, Zsolt MEZRICZKY, Anna MIHÁLY
3D two-photon measurement of single cell and neuronal network activities in vivo. Understanding brain function requires novel imaging methods such as 3D random access point scanning that can simultaneously read out neural activity on both the dendritic and somatic scales. Our 3D AO scanning method can increase measurement speed and signal-to noise ratio by to 6-9 orders of magnitude, but can have one limiting factor: fluorescence information might be lost during brain movement in awake, behaving animals as the amplitude of brain motion is much larger than the diameter of a single excitation spot.
We developed a novel fluorescent imaging technology, which can extend each scanning point to small 3D lines, surface or volume elements, preserving fluorescence information for fast off-line motion correction. Our method effectively eliminates in vivo motion artefacts, allowing fast 3D measurement of over 100 dendritic spines with 3D lines, over 100 somata with squares and cubes, or multiple spiny dendritic segments with surface and volume elements in moving animals. We used this new technology to record activity of pyramidal cells and inhibitory neurons in the moving brain of behaving animals. We revealed a new, broadcasted signalling pathway which activates learning mechanism through the entire neocortex during reward and punishment.
