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3x3D imaging project

Fast two-photon in vivo imaging and stimulation with simultaneous three-dimensional random-access scanning in multiple brain regions 3x3D imaging

We developed and prototyped a novel three-dimensional two-photon laser scanning microscope which can simultaneously image different brain regions in three spatial dimensions (3x3D system). The volumes imaged could exceed cubic millimeters. Such a 3x3D system is essential in understanding distributed brain computation since it allows the simultaneous recording of neuronal activity in multiple, distant brain volumes that are functionally connected. As each site is to be scanned with sub-millisecond temporal resolution in several hundred locations, the connectivity of distant neuronal microcircuits or even neuronal compartments such as dendrites or axons will be followed.
During the years of the 3x3Dimaging project, we have finished the development of the necessary technologies and built the prototype of the 3x3D microscope product. The project started with a kick-off meeting in Budapest on 07/31/2013 and ended on the same location with a review meeting on 01/17/2017. In the third year we encountered various technical challenges and faced unexpected complications when building the 3x3D microscope prototype. To correct these delays, we asked for a six-month non-cost project extension during which we not only completed the 3x3D microscope prototype but also performed the first biological measurements with the device. Many new, unexpected methods and results were achieved and published during the project in high impact journals such as Science, Nature Communications, Neuron, and Nature Neuroscience. The collaborative scientific research embedded in the project proved to be exceptionally fruitful. Our technology will cause long lasting changes to Science, since new principles about neuronal circuit functions across distant brain regions will be discovered. To Technology, since the 3x3D system will enable large brain regions to be scanned in 3D at a cellular resolution. To Society, since the effects of retinal vision restoration in animal models of blindness can be tested in higher visual centers at an unprecedented resolution. To Theory, since theories about distributed brain computations can now be directly tested across multiple brain regions.

Funder: European Comission Research & Innovation FP7

Project period: 2013.06.01.-2016.05.31.

Total cost: 656 440 EUR

EU contribution: 452 550 EUR

Reimbursement rate: 68,94 %


Project Highlights

WP1 Development of new acousto-optical (AO) deflectors for the 3x3D system

WP2 Development of the 3x3D microscope

WP3 Development of ultrafast lasers for deep imaging and efficient optogenetic activation

WP4 Development of control hardware and 3D software

WP5 Neurophysiological and pharmaceutical demonstration in vitro and in vivo

WP6 Demonstration of simultaneous 3D measurements and photo-activation in vivo at depth

WP7 Development of the product prototype

WP8 Management

WP9 Dissemination, Collaboration and Exploitation



Coordinating person: Dr. Balázs Rózsa



Femtonics Kft.

Friedrich Miescher Institute, Basel

Max Delbrück Center for Molecular Medicine

University of Szeged

Budapest University of Technology and Economics


Project details:

Proposal number: 323945

Acronym: 3x3DImaging

Duration of the Project: 36 months

Strategic objective/theme: ICT-2013.9.2