Journal: Advanced Healthcare Materials

Article Title: Environmentally Controlled Microfluidic System Enabling Immune Cell Flow and Activation in an Endothelialised Skin‐On‐Chip

doi: 10.1002/adhm.202400750

Figure Lengend Snippet: Schematic diagram of the microfluidic multi‐well adaptor (MMA) construct. A) Main manifold connecting (a2) external fluidic routing tubes for medium perfusion and allowing light transmission through (a1) the traversing round apertures. B) Biocompatible double‐side adhesive tape layer (142 µm‐thick) with through holes allowing (b2) fluidic routing and (b1) light transmission from (A) to (C). C) High transparency and auto‐fluorescence‐free (188 µm‐thick) COP layer with through holes allowing fluidic routing (c2) from (B) to (D). D) Biocompatible double‐side adhesive tape layer (142 µm‐thick) with patterned microfluidic channels (500 µm‐wide) allowing fluidic routing to the manifold (E) and between different wells (d2). (D) contains through holes for light transmission (d1) from (C) to (E). E) Manifold with through holes (e2) and nozzles (e4) allowing fluidic routing from (A) to a standard 6‐well plate (6MWP). Part (E) presents optical apertures (e1) allowing the transmission of light from a microscope to the biological sample once routed through (A), (B), (C), and (D). Part (E) has also assembled toroidal O‐rings (e5) guaranteeing the sealing, while assembled to the 6MWP, of the overall structure to external factors, such as contamination or gas environment. All parts have some extra features (b3, c3, d3, e3) to allow alignment of the multiple layers and to ease the assembling of the MMA.

Article Snippet: A device with microfluidic channels was specifically designed, developed, and constructed based on patented technologies (Cherry Biotech, US11643632B2, FR3094012B1, and EP3712244A1).

Techniques: Construct, Transmission Assay, Adhesive, Fluorescence, Microscopy

Journal: Advanced Healthcare Materials

Article Title: Environmentally Controlled Microfluidic System Enabling Immune Cell Flow and Activation in an Endothelialised Skin‐On‐Chip

doi: 10.1002/adhm.202400750

Figure Lengend Snippet: Assembly of the complete skin‐on‐chip (SoC) microfluidic device and flow characterization during perfusion. A) Expanded view of the MMA‐6MWP assembly including the MMA, the transwell cell culture insert, containing the RhS, and the 6MWP. B) Complete internal fluidic sealed structure of the myeloid cell‐complemented SoC model. The MMA connects three wells in a series. The direction of flow is indicated with dashed arrows. It is designed to maintain a very low volume of media in the entering well of the 6MWP holding circulating immune cells (“immune cell reservoir”). The second well connected to the previous one contains the RhS (“tissue reservoir”) and is designed to allow the flowed medium to contact only the EC layer at the bottom of the transwell insert. The third well works as a medium collector (“collection reservoir”). Excess medium is collected into an Erlenmeyer flask (“collection flask”). At the end of each experiment, the RhS and the flowed media can be recovered by opening the assembly. Created with BioRender.com. C) Modelled WSS at the transwell membrane: when applying a flow of 150 µL min −1 , WSS values range between 2.95 × 10 −4 Pa and 1.63 × 10 −3 Pa, lower than those reported for human blood vessels in literature. D) Modelled Reynolds number at 1 µm under the transwell membrane when applying a 150 µL min −1 flow. Values range between 3.85 × 10 −6 and 2.36 × 10 −5 (laminarity regime under membrane). E) Heating holder of the assembled SoC MMA‐6WMP. F) Cross‐section of the heating holder that shows the MMA‐6MWP‐holder ensemble. A custom‐made polyamide heater integrated into the base of an aluminum plate warms the MMA‐6MWP ensemble, which has been designed to be compatible with real‐time imaging using a Leica DMi8 Inverted stage. G) Temperature calibration of the system was carried out by placing temperature probes (JTs) in three coaxial regions of three different wells near to the transwell membrane. The results allowed to assess H) zonal and I) mean weighted temperature of the culture medium to guarantee appropriate calibration of the temperatures set by the controlling unit.

Article Snippet: A device with microfluidic channels was specifically designed, developed, and constructed based on patented technologies (Cherry Biotech, US11643632B2, FR3094012B1, and EP3712244A1).

Techniques: Cell Culture, Membrane, Imaging

Journal: Advanced Healthcare Materials

Article Title: Environmentally Controlled Microfluidic System Enabling Immune Cell Flow and Activation in an Endothelialised Skin‐On‐Chip

doi: 10.1002/adhm.202400750

Figure Lengend Snippet: Complete MPS platform prototype (CubiX MVP2C) controlling the gaseous environment (percentages of CO , N , and O ), the perfusion (flow rate), and the heating (temperature) of the myeloid cell‐complemented SoC without the need for an external incubator. A detail of the constructed multi‐well microfluidic adaptor (MMA) is presented in the top‐right of the figure. Medium circulates from the pressurized medium bottle to the “collection flask” via the MMA as depicted by the black dashed arrows. The main components of the CubiX‐MMA‐6MWP‐heater system described in Figures and are labeled in white boxes.

Article Snippet: A device with microfluidic channels was specifically designed, developed, and constructed based on patented technologies (Cherry Biotech, US11643632B2, FR3094012B1, and EP3712244A1).

Techniques: Construct, Labeling

Journal: Micromachines

Article Title: Engineering Microfluidic Organoid-on-a-Chip Platforms

doi: 10.3390/mi10030165

Figure Lengend Snippet: Comparison of advantages and disadvantages of microfluidic chips and other in vitro culture models.

Article Snippet: Although a variety of microfluidic organ-on-a-chip devices with biomimetic compositions, architecture, and functions have been created, there are limited efforts in using reporters and microsensors to monitor different cellular parameters such as growth or metabolism and changes in the culture in the system.

Techniques: Comparison, In Vitro, Cell Culture, Membrane, Concentration Assay, Control

Journal: Micromachines

Article Title: Engineering Microfluidic Organoid-on-a-Chip Platforms

doi: 10.3390/mi10030165

Figure Lengend Snippet: Current organoid-on-a-chip models. ( a ) Brain organoid development in the microchip compartment. The top membrane is coupled to a media reservoir, and the bottom coverslip enables in situ imaging. Wrinkling is caused by nuclear motion and position-dependent nuclear swelling. The differential growth leads to residual stress and wrinkling in the organoid . ( b ) Fluorescence images showing the development of the organoid embedded in Matrigel, and the emergence of wrinkles. Arrows indicate initial wrinkling instability . ( c ) The configuration of the brain organoids-on-a-chip device and the procedures for brain organoids generation on the chip. The EBs formed by hiPSCs were embedded in Matrigel, and the mixtures were infused into the culture channel. The EBs differentiated and self-organized into brain organoids . ( d ) abnormal neurite outgrowth induced by nicotine exposure in the brain organoid-on-a-chip . ( e ) The integrated microfluidic device consisting of modular components including microbioreactors, breadboard, reservoir, bubble trap, physical sensors, and electrochemical biosensors . ( f ) Continual measurements of temperature, pH, and oxygen concentration within the integrated organoid-on-chips. The organoid-on-chip allows in-line automated electrochemical measurements of albumin and GST-α secreted from the hepatic organoids as well as CK-MB from the cardiac organoids. Beating analysis of the cardiac organoids can also be performed . ( g ) Schematic representation of the intestine chip, showing the epithelial (blue) and microvascular (pink) microchannels separated by a porous PDMS membrane sandwiched in-between. The elastic membrane can be extended and retracted by the application of cyclic vacuum to the hollow side chambers. This actuation causes the mechanical deformation of the tissue layers cultured in the chip. ( b ) Procedure to establish the microfluidic co-cultures of the primary human intestinal epithelium and intestinal microvascular endothelium in the intestine chip . ( h ) Generation of hiPSC-derived liver organoids in vitro and the configuration of the liver organoid-on-a-chip system. . Reproduced with permission from [ , , , , ].

Article Snippet: Although a variety of microfluidic organ-on-a-chip devices with biomimetic compositions, architecture, and functions have been created, there are limited efforts in using reporters and microsensors to monitor different cellular parameters such as growth or metabolism and changes in the culture in the system.

Techniques: MicroChIP Assay, Membrane, In Situ, Imaging, Fluorescence, Concentration Assay, Cell Culture, Derivative Assay, In Vitro

Journal: Nature Communications

Article Title: Microfluidics for understanding model organisms

doi: 10.1038/s41467-022-30814-6

Figure Lengend Snippet: Novel microfabrication techniques combined with creativity enable the production of different microfluidic tools that, in turn, facilitate the development of novel experimental paradigms to be used with small multicellular model organisms. Microfluidics empower researchers with tremendous control over sample stimulation while also enabling precise manipulation of samples, and being compatible with high-resolution live imaging. The potential of these microtechnologies is accelerating as they are integrated with additional technologies, become more accessible, and are designed to be multifunctional. Figure created with BioRender.com.

Article Snippet: Fig. 3 Microfluidic systems can deliver stimulations with high precision and at throughputs not previously attainable while making once laborious experiments less demanding. a - i A 3D-printed microfluidic device compatible with a custom-built light-sheet microscope to stimulate zebrafish with precise flow vectors for brain-wide calcium imaging . a - ii A microfluidic device with an array of microposts for analysis of sleep behavior of C. elegans . b A microfluidic system that automatically aligned Drosophila embryos and precisely compressed them using pneumatically actuated deformable sidewalls with simultaneous live imaging . c A microfluidic system capable of trapping hundreds of C. elegans embryos quickly and enabling efficient reagent exchange . d - i A microfluidic system visualized the response of olfactory receptor neurons (ORNs) of Drosophila larva in response to controlled odorant exposure . d - ii A microfluidic device with integrated glass capillaries and a microneedle for chemical injection of Drosophila larvae . e A microfluidic device exposed Drosophila embryo to a thermal gradient along the anterior-posterior axis using two laminar flow streams with different temperatures .

Techniques: Control, Imaging

Journal: Nature Communications

Article Title: Microfluidics for understanding model organisms

doi: 10.1038/s41467-022-30814-6

Figure Lengend Snippet: a-i A microfluidic device with a shape memory alloy actuator immobilized zebrafish and examined the hydrodynamic flow resultant from tail beating . a - ii A worm chamber for visualization and periodic immobilization of C. elegans . b A microfluidic device coupled with a live detection system designed to gently immobilize, orient, and inject zebrafish larvae . c - i A microfluidic system to sort fruit fly samples at high throughput and enrichment ratio . c - ii A non-invasive zebrafish larvae sorting system based on microfluidics that utilized light and acoustics to corral individual samples . All panels are cropped and adapted versions of the originals. Panel c - ii was reproduced from Mani and Chen , with the permission of AIP Publishing.

Article Snippet: Fig. 3 Microfluidic systems can deliver stimulations with high precision and at throughputs not previously attainable while making once laborious experiments less demanding. a - i A 3D-printed microfluidic device compatible with a custom-built light-sheet microscope to stimulate zebrafish with precise flow vectors for brain-wide calcium imaging . a - ii A microfluidic device with an array of microposts for analysis of sleep behavior of C. elegans . b A microfluidic system that automatically aligned Drosophila embryos and precisely compressed them using pneumatically actuated deformable sidewalls with simultaneous live imaging . c A microfluidic system capable of trapping hundreds of C. elegans embryos quickly and enabling efficient reagent exchange . d - i A microfluidic system visualized the response of olfactory receptor neurons (ORNs) of Drosophila larva in response to controlled odorant exposure . d - ii A microfluidic device with integrated glass capillaries and a microneedle for chemical injection of Drosophila larvae . e A microfluidic device exposed Drosophila embryo to a thermal gradient along the anterior-posterior axis using two laminar flow streams with different temperatures .

Techniques: High Throughput Screening Assay

Journal: Nature Communications

Article Title: Microfluidics for understanding model organisms

doi: 10.1038/s41467-022-30814-6

Figure Lengend Snippet: a - i A 3D-printed microfluidic device compatible with a custom-built light-sheet microscope to stimulate zebrafish with precise flow vectors for brain-wide calcium imaging . a - ii A microfluidic device with an array of microposts for analysis of sleep behavior of C. elegans . b A microfluidic system that automatically aligned Drosophila embryos and precisely compressed them using pneumatically actuated deformable sidewalls with simultaneous live imaging . c A microfluidic system capable of trapping hundreds of C. elegans embryos quickly and enabling efficient reagent exchange . d - i A microfluidic system visualized the response of olfactory receptor neurons (ORNs) of Drosophila larva in response to controlled odorant exposure . d - ii A microfluidic device with integrated glass capillaries and a microneedle for chemical injection of Drosophila larvae . e A microfluidic device exposed Drosophila embryo to a thermal gradient along the anterior-posterior axis using two laminar flow streams with different temperatures . All panels are cropped and adapted versions of the originals. Panel c is adapted with permission from Charles et al. , copyright 2020 American Chemical Society.

Article Snippet: Fig. 3 Microfluidic systems can deliver stimulations with high precision and at throughputs not previously attainable while making once laborious experiments less demanding. a - i A 3D-printed microfluidic device compatible with a custom-built light-sheet microscope to stimulate zebrafish with precise flow vectors for brain-wide calcium imaging . a - ii A microfluidic device with an array of microposts for analysis of sleep behavior of C. elegans . b A microfluidic system that automatically aligned Drosophila embryos and precisely compressed them using pneumatically actuated deformable sidewalls with simultaneous live imaging . c A microfluidic system capable of trapping hundreds of C. elegans embryos quickly and enabling efficient reagent exchange . d - i A microfluidic system visualized the response of olfactory receptor neurons (ORNs) of Drosophila larva in response to controlled odorant exposure . d - ii A microfluidic device with integrated glass capillaries and a microneedle for chemical injection of Drosophila larvae . e A microfluidic device exposed Drosophila embryo to a thermal gradient along the anterior-posterior axis using two laminar flow streams with different temperatures .

Techniques: Microscopy, Imaging, Injection