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Biology subjects

Werner, H.

Publications and source records attributed to Werner, H..

3 recordsLinked to original sources

Spatiotemporal control of a multilayered co-axial flow in a 3D printed microchannel with cascaded nozzles

Sculpting and stopping multilayered co-flowing streams is challenging due to inhomogeneous pressure distribution within a fluidic circuit composed of multiple interconnected microchannels having variable flow resistances. Here, we have investigated three different flow control methods to effectively stop a multilayered flow inside a 3D-printed microfluidic channel by bringing the average flow velocity from >100 mm s-1 to below a critical velocity of 200 {micro}m s-1 within a certain delay time tD of [~]2s. Firstly, we 3D printed a sequence of three concentric nozzles ([~]75 {micro}m) embedded serially inside the microchannel ([~]200 {micro}m) using a two-photon polymerization (2PP) method. Secondly, we used the 2PP-based 3D printed device to produce a structured coaxial flow of four streams with individual layer thicknesses of O(10 {micro}m) within the outlet section of the microchannel. Thirdly, we removed the pressure gradient across the fluidic circuit, from > 2 bar to [~]0 bar, to stop the multilayered flow and measured tD to assess the performance of the three stop flow methods. During the stop-flow phase, an inhomogeneous pressure gradient across different inlets resulted in a backflow to inlet channels with lower pressures. In the three stop-flow methods investigated, we systemically managed the fluidic capacitance to minimize a dimensionless backflow index (BFI) value from [~]0.3 (worst case) to [~]0.03 (best case) for a total flow rate ranging from 16.8 {micro}l min-1 to 168 {micro}l min-1. Finally, we have recommended the best stop-flow conditions, which resulted in a minimal delay time of tD [~] 2s and a BFI < 0.05.

bioengineering↗

Fabrication of size-coded amphiphilic particles with a configurable 3D-printed microfluidic device for the formation of particle-templated droplets

Compartmentalizing an aqueous media into numerous nanoliter-scale droplets has substantially improved the performance of amplification assays. Particle-templated droplets or dropicles offer a user-friendly workflow for creating uniform volume compartments upon simple mixing of reagents and particles by using common laboratory apparatus. Amphiphilic shape-coded particles have been demonstrated to spontaneously hold aqueous droplets within hydrophilic cavities for multiplexed diagnostic assays. Here, we have proposed a configurable 3D-printed microfluidic device for the tunable fabrication of amphiphilic size-coded particles. The device was configured with multiple outlet tubings of different diameters and photomasks of variable slit lengths to fabricate a wide range of size-coded particles. We have fabricated >10 unique particle codes using a single reconfigurable device. The cross-sectional profile of the particles was further engineered by tuning the flow rate ratios of precursor streams to vary the inner and outer diameters of the particles and the thicknesses of the inner hydrophilic and outer hydrophobic layers. A range of cavity diameters and particle lengths enabled dropicle volumes of [~]1nL to [~]30nL. The fabricated particles were characterized by their ability to hold uniform droplet volumes and to orient themselves facing upwards or sideways in a well plate based on their aspect ratios.

bioengineering↗

Oligodendrocyte-derived exosomes promote axonal transport and axonal long-term maintenance

Neurons extend long axons that require maintenance and are susceptible to degeneration. Long-term integrity of axons depends on intrinsic mechanisms including axonal transport and extrinsic support from adjacent glial cells. The mechanisms of support provided by myelinating oligodendrocytes to underlying axons are only partly understood. Oligodendrocytes release extracellular vesicles (EVs) with properties of exosomes, which upon delivery to neurons improve neuronal viability in vitro. Here, we show that oligodendroglial exosome secretion is impaired in two mouse mutants exhibiting secondary axonal degeneration due to oligodendrocyte-specific gene defects. Wildtype oligodendroglial exosomes support neurons by improving the metabolic state and promoting axonal transport in nutrient deprived neurons. Mutant oligodendrocytes release less exosomes that share a common signature of underrepresented proteins. Notably, mutant exosomes lack the ability to support nutrient deprived neurons and to promote axonal transport. Together, these findings indicate that glia to neuron exosome transfer promotes neuronal long-term maintenance by facilitating axonal transport, providing a novel mechanistic link between myelin diseases and secondary loss of axonal integrity.

neuroscience↗