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Biology subjects

Serwane, F.

Publications and source records attributed to Serwane, F..

3 recordsLinked to original sources

Scale invariance of mechanical properties in the developing mammalian retina

The computational capabilities of the human central nervous system arise from neuronal architectures. Neuronal tissues are built through force-driven physical remodeling over durations ranging from seconds to days. However, how cell-generated forces accumulate and relax to drive neurogenesis remains unknown due to the difficulty of applying stresses (i) in developing mammalian nervous tissue, (ii) directly within the cellular microenvironment, and (iii) for durations spanning seconds to hours. Previous studies have shown that non-neuronal tissues and cells in 2D culture remodel in scale-free manners. Whether and how this translates to developing neuronal tissues remains an open question. Here, we probed the mechanics of mammalian neuronal tissue on developmental timescales. We accessed developing mammalian neuronal tissue using retinal organoids, stem-cell-derived models of the retina. To probe mechanics at scales relevant for retinal development, we used magnetic droplets as long-term mechanical actuators. We recorded strain responses to applied stresses across four orders of magnitude in time, up to one hour. We found that dynamic creep compliance and tensile moduli follow a power law with an exponent consistent with a material just above the glass transition. This scale-free rheology represents an unprecedented description of nervous tissue mechanics on developmental timescales and opens the door to a biophysical understanding of the emergence of functional neuronal architectures.

bioengineering↗

3D quantification of viral transduction efficiency in living human retinal organoids

The development of therapeutics builds on testing their efficiency in vitro. To optimize gene therapies, for example, fluorescent reporters expressed by treated cells are typically utilized as readouts. Traditionally, their global fluorescence signal has been used as an estimate of transduction efficiency. However, analysis in individual cells within a living 3D tissue remains a challenge. Readout on a single-cell level can be realized via fluorescence-based flow cytometry at the cost of tissue dissociation and loss of spatial information. Complementary, spatial information is accessible via immunofluorescence of fixed samples. Both approaches impede time-dependent studies on the delivery of the vector to the cells. Here, quantitative 3D characterization of viral transduction efficiencies in living retinal organoids is introduced. The approach combines quantified gene delivery efficiency in space and time, leveraging human retinal organoids, engineered adeno-associated virus (AAV) vectors, confocal live imaging, and deep learning-based image segmentation. The integration of these tools in an organoid imaging and analysis pipeline allows quantitative testing of future treatments and other gene delivery methods. It has the potential to guide the development of therapies in biomedical applications.

neuroscience↗

Light-sheet microscopy of network activity in 3D neuronal systems

In vitro systems mimicking brain regions, brain organoids, are revolutionizing the neuroscience field. However, characterization of their electrical activity has remained a challenge as this requires readout at millisecond timescale in 3D at single-neuron resolution. While custom-built microscopes used with genetically encoded sensors are now opening this door, a full 3D characterization of organoid neural activity has not been performed yet, limited by the combined complexity of the optical and the biological system. Here, we introduce an accessible minimalistic light-sheet microscope to the neuroscience community. Designed as an add-on to a standard inverted microscope it can be assembled within one day. In contrast to existing simplistic setups, our platform is suited to record volumetric calcium traces. We successfully extracted 4D calcium traces at high temporal resolution by using a lightweight piezo stage to allow for 5Hz volumetric scanning combined with a processing pipeline for true 3D neuronal trace segmentation. As a proof of principle, we created a 3D connectivity map of a stem cell derived neuron spheroid by imaging its activity. Our fast, low complexity setup empowers researchers to study the formation of neuronal networks in vitro for fundamental and neurodegeneration research.

neuroscience↗