PhD Scientific Days 2018

Budapest, April 19–20, 2018

Novel long-term 3D imaging method for recording neuronal activity and plasticity by acusto-optical microscopy

Pinke, Domonkos

Domonkos Pinke1, Máté Marosi1, Gergely Dobos2, Csaba Csupernyák1, Dominika Nagy1, Gergely Szalay1, Andrius Plauska1, Gergely Katona1, Balázs Rózsa1
1 Institute of Experimental Medicine of Hungarian Academy of Sciences, Budapest
2 Bay Zoltán Foundation for Applied Research, Budapest

Language of the presentation


Text of the abstract


The systematic understanding of brain function requires methods that allow neuronal activity to be recorded at different spatial scales in 3D at a high temporal resolution. Recording techniques are required that collect information from a neuronal population situated in an extensive volume of tissue.
Aims: Technical challenge of two-photon imaging in long-term is the stability of cell recordings. Lateral and Z displacement can be managed with 2D shift scanning and Z-focusing, however, effects of XYZ rotation and tissue deformation remain unsolved.


Here we present a novel fast 3D volumetric imaging method, suitable for long-term in vivo tracking of neuronal activity in mouse cortex by acusto-optical two-photon microscopy. We could precisely identify and record from the center plane of cell bodies and track the visually evoked neuronal activity. On the first day of the experiment after a control Z-stack acquisition in near-cubic-millimeter scan range, 3D drifting acusto-optical volumetric imaging (50x50x50µm Multi-cube scans) were recorded around the selected cell bodies. On the next imaging sessions the recording coordinates were re-loaded to the acquisition software and the recording sites were roughly identified by the vascular architecture. Then new Multi-cube scans were acquired and used for fine alignment in a 3D volume.


With the Multi-cube scanning method we are able to locate and record Ca2+ activity (with GCaMP6f) of the same neuronal ensemble during our 10-20 days long protocol including baseline, training and post learning imaging sessions of up to 200 cells. Our results show this method is significantly more accurate than a conventional Z-stack ROI selection.


We demonstrate a method, that allows us to record long term neuronal plasticity from the same cell population in a near-cubic millimeter 3D volume, up to 2 months. For this reason, the Multi-cube scanning method is suitable for long term imaging behavior protocols.

Data of the presenter

Doctoral School: Szentágothai János Doctoral School of Neuroscience
Program: Functional Neuroscience
Supervisor: Balázs Rózsa
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