Devices and systems for molecular barcoding of nucleic acid targets in single cells
US-2016289669-A1 · Oct 6, 2016 · US
US2018200710A1 · US · A1
| Field | Value |
|---|---|
| Publication number | US-2018200710-A1 |
| Application number | US-201815868811-A |
| Country | US |
| Kind code | A1 |
| Filing date | Jan 11, 2018 |
| Priority date | Jan 13, 2017 |
| Publication date | Jul 19, 2018 |
| Grant date | — |
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Disclosed herein are methods, devices, and systems for efficient loading and retrieval of particles. In some embodiments, a fluidic channel of a flowcell comprises a ceiling, a first sidewall, and a bottom, wherein the contact angle of the ceiling is at least 10 degrees smaller than the contact angle of the first sidewall, and wherein the bottom of the fluidic channel comprises a substrate that comprises a plurality of microwells.
Opening claim text (preview).
What is claimed is: 1 . A device comprising: a flowcell comprising a fluidic channel, an inlet port, and an outlet port, wherein the fluidic channel comprises a ceiling, a first sidewall, and a fluidic channel bottom, wherein the ceiling and the first sidewall form a ceiling-first sidewall edge, and wherein the bottom and the first sidewall for a bottom-first sidewall edge, wherein the contact angle of the ceiling is at least 10 degrees smaller than the contact angle of the first sidewall, wherein the bottom of the fluidic channel comprises a substrate which comprises a plurality of microwells, and wherein the inlet port and the outlet port are in fluid communication with the flowcell via the fluidic channel. 2 . The device of claim 1 , wherein the fluidic channel has a non-circular cross section. 3 . The device of claim 1 , wherein the fluidic channel comprises a second sidewall, wherein the ceiling and the second sidewall form a ceiling-second sidewall edge, and wherein the bottom and the second sidewall for a bottom-second sidewall edge. 4 . The device of claim 1 , wherein the fluidic channel has a width of at least 2 mm. 5 . The device of claim 1 , wherein the fluidic channel is at least 0.8 mm in height. 6 . The device of claim 1 , wherein the first sidewall has a positive draft angle of 1-15 degrees. 7 . The device of claim 1 , wherein the flowcell comprises silicon, fused-silica, glass, polydimethylsiloxane (PDMS), polymethylmethacrylate (PMMA), polycarbonate (PC), polypropylene (PP), polyethylene (PE), high density polyethylene (HDPE), polyimide, cyclic olefin polymers (COP), cyclic olefin copolymers (COC), polyethylene terephthalate (PET), epoxy resin, metal, or a combination thereof. 8 . The device of claim 1 , wherein the ceiling is hydrophilic. 9 . The device of claim 1 , wherein the contact angle of the ceiling is 0-80 degrees. 10 . The device of claim 1 , wherein the contact angle of the first sidewall is about 88 degrees. 11 . The device of claim 1 , wherein the contact angle of the ceiling is sufficiently smaller than the contact angle of the first sidewall to enable non-laminar flow within the fluidic channel. 12 . The device claim 11 , wherein the non-laminar flow within the fluidic channel enables agitation of a particle on the surface of the substrate by a flow within the fluidic channel, and wherein a speed of the flow at a boundary between the flow and the bottom is non-zero. 13 . The device of claim 11 , wherein the non-laminar flow is approximately plug flow, and wherein the relative flow velocity of a flow across a cross section of the fluidic channel is approximately constant. 14 . The device of claim 11 , wherein the non-laminar flow is plug flow, and wherein the relative flow velocity of a flow across a cross section of the fluidic channel is constant. 15 . The device of claim 14 , wherein the plug flow is approximately horizontal plug flow. 16 . The device of claim 15 , wherein the horizontal plug flow is capillary aided horizontal plug flow. 17 . The device of claim 14 , wherein the plug flow does not depend on buoyancy of a gas, or wherein the plug flow does not depend on tilting of the device. 18 . The device of claim 14 , wherein the plug flow is at a buffer-gas interface. 19 . The device of claim 1 , wherein the ceiling comprises a hydrophilic coating. 20 . The device of claim 19 , wherein the hydrophilic coating comprises polyethylene glycol (PEG), poly-Hema, pluronic acid F68, pluronic acid F108, pluronic acid F127, polysorbate 20, silicon dioxide (SiO 2 ), silicon nitride, or any combination thereof. 21 . The device of claim 20 , wherein the hydrophilic coating comprises silicon dioxide (SiO 2 ), silicon nitride, or any combination thereof. 22 . The device of claim 19 , wherein the contact angle of the hydrophilic coating is at most 23 degrees. 23 . The device of claim 19 , wherein the ceiling is coated with the hydrophilic coating by sputtering, thermal growth, adsorption, covalent binding, or any combination thereof. 24 . The device of claim 19 , wherein the hydrophilic coating is offset from the ceiling-first sidewall edge. 25 . The device of claim 24 , wherein the hydrophilic coating is offset from the ceiling-first sidewall edge by 100-1000 micrometers. 26 . The device of claim 24 , wherein the hydrophilic coating is offset from the ceiling-first sidewall edge by 1%-25% of the width of the ceiling. 27 . The device of claim 1 , comprising a plurality of particles residing in the plurality of microwells of the substrate, and wherein at least 25% of the plurality of microwells each contains a single particle. 28 . A method for sample loading, comprising (a) providing a device that comprises: a flowcell comprising a fluidic channel, an inlet port, and an outlet port, wherein the fluidic channel comprises a ceiling, a first sidewall, and a bottom, wherein the ceiling and the first sidewall forms a ceiling-first sidewall edge, and wherein the bottom and the first sidewall forms a bottom-first sidewall edge, wherein the contact angle of the ceiling is at least 10 degrees smaller than the contact angle of the first sidewall, wherein the bottom comprises a substrate which comprises a plurality of microwells, and wherein the inlet port and the outlet port are in fluid communication with the flowcell via the fluidic channel; (b) introducing a gas into the fluidic channel via the inlet port; and (c) introducing a first sample comprising a first plurality of particles into the fluidic channel via the inlet port, whereby at least 25% of the plurality of microwells each contains a single particle of the first plurality of particles. 29 . The method of claim 28 , comprising: (d) introducing the gas or another gas into the fluidic channel via the inlet port after step (c); and (e) introducing a second sample into the fluidic channel via the inlet port after introducing the first sample into the fluidic channel, wherein the second sample comprises a second plurality of particles, and whereby at least 25% of the plurality of microwells each contains a single particle of the second plurality of particles. 30 . The method of claim 29 , comprising: prior to introducing the gas into the fluidic channel via the inlet port in (d), introducing a first buffer into the fluidic channel via the inlet port to remove the first plurality of particles that are on the bottom and are not contained in any of the plurality of microwells, or after introducing the second sample into the fluidic channel via the inlet port in (e), introducing a second buffer into the fluidic channel via the inlet port to remove the second plurality of particles that are on the bottom and are not contained in any of the plurality of microwells.
Handling flowable solids, e.g. microscopic beads, cells, particles · CPC title
by measuring electrical or magnetic effects · CPC title
having a very large number of wells, microfabricated wells · CPC title
with fluid transport, e.g. in multi-compartment structures · CPC title
specially adapted for droplet or plug flow, e.g. digital microfluidics · CPC title
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