Methods for encapsulating and assaying cells
Inventors
Kurz, Volker L. S. • McEwen, Jason M. • Mobilia, Kellen C. • MASTROIANNI, Alexander J. • CARDIEL RIVERA, Joshua J.
Assignees
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Publication Number
US-11612890-B2
Publication Date
2023-03-28
Expiration Date
Abstract
In biosciences and related fields, it can be useful to study cells in isolation so that cells having unique and desirable properties can be identified within a heterogenous mixture of cells. Processes and methods disclosed herein provide for encapsulating cells within a microfluidic device and assaying the encapsulated cells. Encapsulation can, among other benefits, facilitate analyses of cells that generate secretions of interest which would otherwise rapidly diffuse away or mix with the secretions of other cells.
Core Innovation
The invention relates to microfluidic devices and processes that reversibly encapsulate cells in a channel connected to a plurality of chambers. Each chamber includes an opening that fluidically connects the chamber to the channel, and surfaces forming the channel proximal to the opening and/or surfaces forming each chamber proximal to the channel include a hydrophobic coating. The enclosure is filled with a first aqueous medium, and cells are disposed in respective chambers so that different cells reside in different chambers.
A water immiscible fluidic medium is then flowed into the channel to displace substantially all of the first aqueous medium in the channel without substantially displacing the first aqueous medium in the chambers. This displacement reversibly encapsulates the cells in their respective chambers and maintains compartmentalization of aqueous media by using the hydrophobic coating and the water immiscible fluidic medium.
In related embodiments, the device enables monitoring and assaying activities of cells that remain encapsulated in separate chambers. The assaying includes monitoring an activity of the first and second cells encapsulated in their respective first and second chambers, and may include flowing a third aqueous medium comprising assay reagent, lytic reagent, or export buffer.
Claims Coverage
Two independent claims are identified. The claims cover reversible encapsulation of cells in chambers connected to a channel by selectively displacing a first aqueous medium from the channel using a water immiscible fluidic medium, while retaining aqueous medium in the chambers, with hydrophobic surface coatings near the openings. One independent claim further requires monitoring an activity for assaying purposes.
Reversible encapsulation by channel displacement while retaining chamber aqueous medium
A process for encapsulating cells in a microfluidic device having an enclosure comprising a channel and a plurality of chambers, wherein the chamber-channel enclosure is filled with a first aqueous medium, cells are disposed in respective chambers, and a water immiscible fluidic medium is flowed into the channel, displacing substantially all of the first aqueous medium in the channel without substantially displacing the first aqueous medium in the chambers, thereby reversibly encapsulating the cells in their respective chambers.
Hydrophobic coating proximal to chamber openings
At least a portion of surfaces forming the channel proximal to the opening to each chamber and/or at least a portion of surfaces forming each chamber proximal to the channel comprises a hydrophobic coating as part of the encapsulation process.
Assaying encapsulated cells by monitoring activity after reversible encapsulation
A process of assaying encapsulated cells in a microfluidic device comprising a channel and a plurality of chambers, filled with a first aqueous medium, with cells disposed in respective chambers, wherein flowing a water immiscible fluidic medium into the channel displaces substantially all of the first aqueous medium in the channel without substantially displacing the first aqueous medium in the chambers, thereby reversibly encapsulating the cells in their respective chambers, and monitoring an activity of the cells encapsulated in the respective chambers.
Selective de-encapsulation and third aqueous medium
The process further selects one or more chambers having a cell disposed therein; removes the encapsulation layer at the opening to the channel of the selected chamber(s) to generate a de-encapsulated chamber; and flows a third aqueous medium into the channel, where the third medium comprises an assay reagent, a lytic reagent, or an export buffer.
The claims collectively cover reversible encapsulation of cells in separate chambers connected to a channel by hydrophobic surface coatings near chamber openings and by selectively displacing aqueous medium from the channel using a water immiscible fluidic medium. The assaying claim extends this framework to monitoring cell activity while the cells remain reversibly encapsulated in their respective chambers.
Stated Advantages
Enables monitoring an activity of cells encapsulated in respective chambers after reversible encapsulation.
Allows displacement of aqueous medium in the channel without substantially displacing aqueous medium in the chambers, thereby reversibly encapsulating cells in their respective chambers.
Documented Applications
Assaying encapsulated cells in a microfluidic device having a channel connected to a plurality of chambers, including monitoring activity of cells in respective chambers.
Assaying encapsulated cells in separate microfluidic chambers by monitoring an activity of cells after reversible encapsulation.
Providing assay workflows that can include flowing a third aqueous medium comprising assay reagent, lytic reagent, or export buffer for downstream use as part of the microfluidic assay process.
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