Controlling the force and the position of acoustic traps with a tunable acoustofluidic chip: Application to spheroid manipulations.

A multi-node acoustofluidic chip working on a broadband spectrum and beyond the resonance is designed for cell manipulations. A simple one-dimensional (1D) multi-layer model is used to describe the stationary standing waves generated inside a cavity. The transmissions and reflections of the acoustic wave through the different layers and interfaces lead to the creation of pressure nodes away from the resonance condition. A transparent cavity and a broadband ultrasonic transducer allow the measurement of the acoustic energy over a wide frequency range using particle image velocimetry measurements and the relation between acoustic energy and the particles velocity. The automation of the setup allows the acquisition over a large spectrum with a high frequency definition. The results show a wide continuous operating range for the acoustofluidic chip, which compares well with the 1D model. The variation of the acoustic radiation force when varying the frequency can be compensated to ensure a constant amplitude for the ARF. This approach is finally applied to mesenchymal stem cell (MCS) spheroids cultured in acoustic levitation. The MSC spheroids can be moved and merged just by varying the acoustic frequency. This approach opens the path to various acoustic manipulations and to complex 3D tissue engineering in acoustic levitation.

Pharmaco-fUS: Quantification of pharmacologically-induced dynamic changes in brain perfusion and connectivity by functional ultrasound imaging in awake mice.

There is a critical need for reliable quantitative biomarkers to assess functional brain alterations in mouse models of neuropsychiatric diseases, but current imaging methods measuring drug effects through the neurovascular coupling, face issues including poor sensitivity, drug-induced changes in global brain perfusion and the effects of anesthesia. Here we demonstrate the proof-of-concept of a minimally-invasive fUS imaging approach to detect the acute cholinergic modulatory effects of Scopolamine (ScoP) on functional brain connectivity in awake and behaving mice, through the intact skull. A machine-learning algorithm constructed an ad-hoc pharmacological score from the ScoP-induced changes in connectivity patterns of five mice. The discrimination model shows important ScoP-induced increase of the hippocampo-cortical connectivity. The pharmacological score led to robust discrimination of ScoP treatment from baseline in an independent dataset and showed, in another independent group, dose-dependent specific effects of central cholinergic modulation of functional connectivity, independent from global brain perfusion changes. In conclusion, we introduce pharmaco-fUS as a simple, robust, specific and sensitive modality to monitor drug effects on perfusion and functional connectivity in the awake mouse brain.

Influence of microgravity on spontaneous calcium activity of primary hippocampal neurons grown in microfluidic chips.

The influence of variations of gravity, either hypergravity or microgravity, on the brain of astronauts is a major concern for long journeys in space, to the Moon or to Mars, or simply long-duration missions on the ISS (International Space Station). Monitoring brain activity, before and after ISS missions already demonstrated important and long term effects on the brains of astronauts. In this study, we focus on the influence of gravity variations at the cellular level on primary hippocampal neurons. A dedicated setup has been designed and built to perform live calcium imaging during parabolic flights. During a CNES (Centre National d'Etudes Spatiales) parabolic flight campaign, we were able to observe and monitor the calcium activity of 2D networks of neurons inside microfluidic devices during gravity changes over different parabolas. Our preliminary results clearly indicate a modification of the calcium activity associated to variations of gravity.