Microfluidic device for enabling fluidic isolation among interconnected compartments within the apparatus and methods relating to same
View Patent ↗Embodiments of the invention are directed to a device that combines microfabrication, microfluidic, and surface micropatterning techniques to create a multi-compartment neuronal culturing device that has application across a number of different neuroscience uses. Devices configured in accordance with the invention allow directed growth of neurites and isolation of neurites from their cell bodies. The device can use hydrostatic pressure to isolate insults to one compartment and, thus, expose localized areas of neurons to insults. Due to the high resistance of the microgrooves for fluid transport, insults are contained in the neuritic compartment without appreciable leakage into the somal compartment for a certain period of time (e.g., over 15 h).
1. A multi-compartment microfluidic device for enabling fluidic isolation among interconnected compartments within the device comprising:
a substrate coupled with an optically transparent device;
said substrate comprising a micropatterned cell-adherent coating configured to direct cell attachment;
said optically transparent device comprising a first microfluidic region having a first plurality of entry reservoirs for accepting or extracting a first volume of fluid;
said optically transparent device further comprising a second microfluidic region, said second microfluidic region having a second plurality of entry reservoirs for accepting or extracting a second volume of fluid that is less than said first volume of fluid to create hydrostatic pressure;
a barrier region that couples said first microfluidic region with said second microfluidic region in a way that enables a biological specimen to simultaneously extend across said first microfluidic region, said barrier region and said second microfluidic region; and,
said barrier region comprising a plurality of microgrooves having a width and height that enables said second volume of fluid to be fluidically isolated from said first volume of fluid via said hydrostatic pressure maintained via said at least one embedded microgroove wherein said first microfluidic region, said second microfluidic region, said first plurality of entry reservoirs, said second plurality of entry reservoirs and said barrier region are fabricated into said optically transparent device.
2. The multi-compartment microfluidic device of claim 1 wherein said first microfluidic region and said second microfluidic region are dispose parallel to one another and coupled with said barrier region.
3. The multi-compartment microfluidic device of claim 1 wherein said at barrier region comprises a length of not less than 50 μm.
4. The multi-compartment device of claim 1 wherein at least one of said plurality of microgrooves comprises dimensions less than 10 μm in height.
5. The multi-compartment microfluidic device of claim 1 wherein said biological specimen comprises a cellular structure.
6. The multi-compartment microfluidic device of claim 5 wherein said first volume of fluid is applied to a cell body domain of said cellular structure and said second volume of fluid is applied to a cellular extension or outgrowth domain of said cellular structure.
7. The multi-compartment microfluidic device of claim 6 wherein said cellular extension or outgrowth domain comprises pseudopod or lamellipodium.
8. The multi-compartment microfluidic device of claim 5 wherein said cellular structure comprises a nerve cell.
9. The multi-compartment microfluidic device of claim 8 wherein said first volume of fluid is applied to a somal domain of said nerve cell and said second volume of fluid is applied to an neuritic region of said nerve cell.
10. The multi-compartment microfluidic device of claim 9 wherein said somal domain comprises a nerve cell body.
11. The multi-compartment microfluidic device of claim 9 wherein said neuritic region comprises an axonal domain.
12. The multi-compartment microfluidic device of claim 8 wherein synapses of said nerve cell are isolated in said second microfluidic region.
13. The multi-compartment microfluidic device of claim 1 wherein said cell-adherent coating comprises any one or more selected from the group consisting of polylysine, laminin, collagen, fibronectin, integrin, polyamine, and polyornithine.
14. A method for enabling fluidic isolation among interconnected compartments within a multi-compartment microfluidic device comprising:
forming a micropatterned cell-adherent coating configured to direct cell attachment onto a substrate;
coupling said substrate with an optically transparent device;
forming into said optically transparent device a first microfluidic region having a first plurality of entry reservoirs for accepting or extracting a first volume of fluid;
forming into said optically transparent device a second microfluidic region, said second microfluidic region having a second plurality of entry reservoirs for accepting or extracting a second volume of fluid that is less than said first volume of fluid to create hydrostatic pressure;
forming into said optically transparent device a barrier region that couples said first microfluidic region with said second microfluidic region in a way that enables a biological specimen to simultaneously extend across said first microfluidic region, said barrier region and said second microfluidic region; and,
isolating fluidically said first volume of fluid from said second volume of fluid using said barrier region comprising a plurality of microgrooves having a width and height that enables said second volume of fluid to be fluidically isolated from said first volume of fluid via said hydrostatic pressure maintained via said at least one embedded microgroove wherein said first microfluidic region, said second microfluidic region, said first plurality of entry reservoirs, said second plurality of entry reservoirs and said barrier region are fabricated into said optically transparent device.