Freezing and archiving cells on a microfluidic device
Inventors
White, Mark P. • Chapman, Kevin T. • McFarland, Andrew W. • Hobbs, Eric D. • Lowe, JR., Randall D.
Assignees
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Publication Number
US-12102083-B2
Publication Date
2024-10-01
Expiration Date
Abstract
A method of processing and storing biological cells includes introducing a flowable medium into a microfluidic device, the flowable medium including biological cells; sequestering one or more biological cells from the flowable medium in one or more isolation regions of the microfluidic device; and freezing the microfluidic device including the one or more biological cells sequestered therein.
Core Innovation
A method of processing and storing biological cells introduces a flowable medium including biological cells into a microfluidic device. The microfluidic device includes a base with an electrode activation substrate and a microfluidic circuit structure with a flow region and one or more isolation chambers, where each isolation chamber includes an isolation region and a connection region with a proximal opening into the flow region and a distal opening into the respective isolation region. The isolation region is an unswept region having a single opening into the respective connection region, and the device further comprises a cover.
The method sequesters one or more biological cells from the flowable medium in one or more isolation regions of the microfluidic device. The sequestering uses the microfluidic circuit structure having isolation chambers with unswept isolation regions and connection regions to isolate the biological cells from the flow region. After sequestering, the method freezes the microfluidic device including the one or more biological cells sequestered therein.
Freezing can be carried out with staged cooling that includes cooling to a near-freezing temperature and then cooling to subzero temperatures. The workflow can also include creating an inventory that includes at least an identity and an isolation region location for each sequestered biological cell, and storing the inventory in memory associated with the microfluidic device. In some workflows, a cell preservation reagent is introduced prior to freezing.
Claims Coverage
The claim coverage centers on a method that combines a specific microfluidic geometry with unswept isolation regions in isolation chambers connected to a flow region and processing steps that sequester biological cells in the isolation regions and freeze the device. The independent claim is supported by dependent inventive features including staged subzero freezing, isolation region volume and connection region geometry constraints, pre-freeze cell preservation reagent introduction, and inventory creation stored in device-associated memory.
Microfluidic device with unswept isolation chambers and connection regions
A microfluidic device comprising a base with an electrode activation substrate and a microfluidic circuit structure disposed thereupon, the microfluidic circuit structure comprising a flow region and one or more isolation chambers, wherein each isolation chamber comprises an isolation region and a connection region, the connection region having a proximal opening into the flow region and a distal opening into the respective isolation region, and wherein the isolation region is an unswept region having a single opening into the respective connection region, the microfluidic device further comprising a cover.
Sequestering cells in unswept isolation regions
Sequestering one or more biological cells from the flowable medium in one or more isolation regions of the microfluidic device.
Freezing the microfluidic device with sequestered cells
Freezing the microfluidic device including the one or more biological cells sequestered therein.
Staged subzero freezing schedule
Freezing the microfluidic device by first cooling it to near-freezing temperature and then further cooling it to a subzero temperature.
Isolation region volume range
Each isolation region has a volume between 1.5×10^5 and 1.5×10^6 cubic microns.
Connection region length-to-width ratio at the proximal opening
The ratio of the connection region length (L_con) to the connection region width (W_con) at the proximal opening into the flow region is at least 1.0 for each isolation chamber.
Pre-freeze cell preservation reagent introduction
Introducing a cell preservation reagent into the microfluidic device before freezing it.
Inventory creation and device-associated memory storage
Creating an inventory including at least an identity and isolation region location for each of the one or more sequestered biological cells, and storing the inventory in a memory associated with the microfluidic device.
The claim set is directed to processing and storing biological cells in a microfluidic device using unswept isolation regions within isolation chambers connected to a flow region, followed by sequestering and freezing of the device. Dependent features further define the chamber geometry and capacity, staged cooling to subzero temperatures, pre-freeze cell preservation reagent introduction, and recording cell identity and isolation-region location in device-associated memory.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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