Minimally invasive endoscopes for neuronal activity monImaging-assisted single-cell specific activity monitoring and optogenetic stimulation of deep brain structures in motile and awaken animal models

Project Information

WOKEGATE

Grant agreement ID: 101069245

DOI

10.3030/101069245

Project closed

EC signature date 10 June 2022

Start date 1 November 2022

End date 30 April 2024

Funded under

• European Research Council (ERC)

Total cost

No data

EU contribution

€ 150 000,00

Coordinated by

LEIBNIZ-INSTITUT FUER PHOTONISCHE TECHNOLOGIEN E.V.
Germany

Periodic Reporting for period 1 - WOKEGATE (Minimally invasive endoscopes for neuronal activity monImaging-assisted single-cell specific activity monitoring and optogenetic stimulation of deep brain structures in motile and awaken animal models)

Reporting period: 2022-11-01 to 2024-04-30

Summary of the context and overall objectives of the project

The WOKEGATE project aimed to empower the discipline of in-vivo neuroscience imaging by developing the technologies to precisely and minimally invasively locate, observe, and manipulate individual neurons in deep brain regions of awake, freely moving animal models. This ambition has been integrated into the already existing translational pathway via the start-up endeavour DeepEn. This ambitious goal was achieved through several key work packages, each contributing to significant advancements in the field of in-vivo brain imaging.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

The core technological development was the creation of a stand-alone module utilizing the M3CF fibre probe in two innovative regimes. The first regime was the High-Resolution Holographic Multimode Fibre Imaging Regime, which is resilient to non-bending and efficiently navigates and targets specific brain structures. The second regime, the Bending-Resilient Multicore Regime, enables the addressing of individual neurons even under fibre deformation. Additionally, a detachable fibre connector and a compact reconnection tool were developed, enhancing the module's usability and flexibility.
Based on the feedback and applicability studies from subsequent work packages, the project also explored the potential of commercially available multimode optical fibres that exhibit bending resilience. This exploration identified fibres capable of maintaining imaging performance even under extreme bending conditions, offering a much simpler alternative to the M3CF. This alternative approach promises high-resolution imaging and neuron activity monitoring in freely moving animal models, albeit within a limited range of free motion space.
The stand-alone module was validated at the German Centre for Neurodegenerative Diseases, University of Magdeburg. The validation involved acute experiments on head-fixed anesthetized mice, successfully utilizing the high-resolution imaging modality to target Purkinje cells in the cerebellum. Recording neuronal activity with the bending-resilient regime encountered a challenge related to low collection efficiency. The approach can still be suitable when using bright fluorescent proteins but would require modifications in future M3CF to allow the use of the most prominent fluorophores, including the greatly desired voltage indicators.
These efforts culminated in the publication "Advancing the path to in-vivo imaging in freely moving mice via multimode-multicore fibre based holographic endoscopy" by Y. Du et al. in Neurophotonics, published in 2024.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

A thorough market and customer analysis was conducted to evaluate the commercial potential of WOKEGATE technology. Key advantages identified include minimal invasiveness, as WOKEGATE's technology causes significantly less damage compared to existing techniques, and high-resolution in-vivo imaging, capable of imaging deep brain structures like the brain stem and amygdala, which are unreachable by other methods. The estimated pricing strategy places the technology competitively between miniscopes and multiphoton microscopes, with a high potential for sustainable sales growth. Market analysis projects an initial market volume of €800 million to €1 billion, with recurring annual demand for probes estimated at €50 million to €75 million.
The contracted research transfer mentor collaborated with neuroscience partners and the DeepEn start-up team, gathering feedback and refining the value proposition. The partnership has led to several key outcomes. The M3CF patent is in the PCT stage, with exclusive licensing to DeepEn. Further findings are under internal disclosure and patent application development. Flexible holographic endoscopy has been incorporated into DeepEn's business plan, with the first product launch planned for 2025.
The DeepEn start-up has been incorporated as a limited company (GmBH) during the execution of the project.
To fund further development, a joint proposal for the EIC Transition is being prepared. Alternative European and German national funding options have been identified. The strategy includes scaling production and preparing for international expansion by 2027, with continued R&D efforts for a second-generation product.
The WOKEGATE project has made significant strides in developing minimally invasive, high-resolution in-vivo brain imaging technologies. Through technological innovation, rigorous validation, market analysis, and strategic collaborations, the project has laid a strong foundation for future advancements and commercialisation in neuroscience research.