Precise delivery of components into fluids
Inventors
Spuhler, Philipp • Li, Sixing • Prosen, Dennis
Assignees
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Publication Number
US-11976269-B2
Publication Date
2024-05-07
Expiration Date
Abstract
Disclosed herein include systems, apparatuses, devices, and methods for introducing one or more components into a fluid. A first fluid and a second fluid can be co-injected into a fluidic channel of a flow cell. In some embodiments, the first fluid and a second fluid are immiscible (e.g. an aqueous buffer and a non-aqueous liquid). In some embodiments, the second fluid is less dense than the first fluid.
Core Innovation
The invention relates to a method for introducing one or more components to contents of microwells by controlled fluid delivery in a fluidic channel having a ceiling, a sidewall, and a bottom with a plurality of microwells. A first fluid is introduced so that the fluidic channel and each microwell comprise the first fluid, and a first displacement fluid then displaces the first fluid from the fluidic channel at a first flow rate.
Subsequently, a second fluid is introduced at a second flow rate while in contact with a second displacement fluid that is immiscible to the second fluid, such that the second fluid is immediately followed by and in contact with the second displacement fluid and the second fluid and the second displacement fluid have the same flow rate. One or more components of the second fluid enter the content in the microwell when the second fluid comes into contact with the content for a first duration.
The second displacement fluid displaces the second fluid from the fluidic channel and/or seals the content of the microwell, reducing cross-talk via diffusion. The disclosed implementations include flowcell priming and sequential co-injection/co-dispensing in microwell arrays using immiscible fluids such as aqueous liquids, non-aqueous liquids, and/or gas, with defined temporal contact such that components diffuse into microwell contents while a second fluid seals the microwells.
Claims Coverage
The independent claim coverage centers on a sequence of fluid introductions and displacements in a microwell-containing fluidic channel. Three inventive features are identified from the provided text.
Microwell channel with priming fluid and first displacement
Introducing a first fluid into a fluidic channel comprising a ceiling, a first sidewall, and a bottom, wherein the bottom comprises a plurality of microwells, whereby the fluidic channel and each microwell comprise the first fluid; introducing a first displacement fluid into the fluidic channel to displace the first fluid from the fluidic channel at a first flow rate.
Immiscible second fluid/displacement-fluid pair with equal flow rate
Introducing a second fluid into the fluidic channel at a second flow rate while in contact with a second displacement fluid that is immiscible to the second fluid such that the second fluid is immediately followed by and in contact with the second displacement fluid and the second fluid and the second displacement fluid have the same flow rate.
Displacement and/or sealing of microwell contents
Wherein the second displacement fluid displaces the second fluid from the fluidic channel and/or seals the content of the microwell.
Overall, the claims center on delivering components to microwell contents using an immiscible second displacement-fluid pair that is sequenced to create a defined contact duration for component entry, while the second displacement fluid displaces and/or seals the microwell contents to reduce undesired diffusion and cross-talk.
Stated Advantages
Reduced cross-talk via sealing.
Reduced cross-talk via diffusion of components by sealing microwells after defined contact.
Higher-density microwell arrays enabled.
Documented Applications
Single-cell nucleic acid quantification including downstream molecular barcoding/digital counting, consistent with barcodes and molecular labels for quantification.
Controlled delivery for reaction initiation, including lysis and downstream reactions such as reverse transcription/extension/PCR.
Effects dosing studies with introduced components and dose-dependent phenotypic effects [procedural detail omitted for safety].
Microwell-array assays described as supporting Rhapsody assays.
An example with CHAPS lysis buffer and oil sealing containing calcein and larger molecules within microwells to support reduced molecular leakage/cross-talk.
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