Methods of using modular assay support devices
Inventors
Cox, David Maurice • Katiraee, Layla
Assignees
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Publication Number
US-12566114-B2
Publication Date
2026-03-03
Expiration Date
Abstract
A substrate holder includes a base configured to receive a substrate; a cover configured to mateably engage with the base, the cover defining an opening formed by inner sidewalls; and a removable insert defining a surface, the removable insert being configured to be received within the opening of the cover. The removable insert includes a gasket; a projection coupled to the gasket; and at least two insert tabs extending from opposite sides of the removable insert, each insert tab being configured to engage with at least one of the inner sidewalls forming the opening of the cover.
Core Innovation
The invention provides modular substrate support devices for spatial bioassays. A substrate is mounted on a base of a substrate holder, and the substrate includes a sample region supporting at least one sample. A cover is releasably locked to the base so that a well is formed around the sample region, and the cover includes an opening arranged within a perimeter of the cover.
Releasably locking the cover to the base is achieved by releasably locking a first cover tab to a first base tab and a second cover tab to a second base tab. This tab-based engagement forms and maintains the well around the sample region while enabling reagent addition through the opening in the cover. The device architecture supports alignment and repositioning of gaskets and ports over different sample regions, including an unlocked state with a gap to adjust insert and gasket placement without disturbing samples, and a releasably locked state achieved by movable and non-movable tabs.
The well and gasket configuration provide vapor-/fluid-tight sealing around the sample region, which is stated to improve sealing against cross-contamination and enable handling of samples in spatial bioassay workflows. The system is also described as adaptable for integration with heating devices and for reducing condensation. The approach is tied to method workflows in which, after releasably locking the cover to the base, a reagent is contacted with the sample by adding the reagent to the well through the opening arranged within the perimeter of the cover.
Dependent embodiments include permeabilizing at least a portion of the sample using a permeabilization agent included in the reagent, followed by spatial analysis of analytes. Further embodiments describe nucleic-acid workflows that include reverse transcription, second-strand cDNA synthesis, and cDNA library preparation, and embodiments that specify glass slide substrates with sample regions including spatially barcoded capture probes.
Claims Coverage
The document includes two independent method claims. Across these claims, the inventive coverage is centered on tab-based releasable locking of a cover to a base to form a well around a sample region, and on contacting the sample with a reagent by adding the reagent to the well through an opening in the cover.
Tab-based cover-to-base releasable locking to form a well
Releasably locking a cover of the substrate holder to the base by releasably locking a first cover tab to a first base tab and a second cover tab to a second base tab thereby forming a well around the sample region.
Reagent addition through a cover opening into the well
After releasably locking the cover to the base, contacting the at least one sample with a reagent by adding the reagent to the well through an opening arranged within the perimeter of the cover.
Well-forming base geometry with laterally extending tabs and side walls
A base comprising a bottom portion, a first side wall extending from the first end of the bottom portion, a second side wall extending from the second end of the bottom portion, and a first base tab and a second base tab extending laterally outward from outer surfaces of the first and second side walls, respectively, with the second side wall arranged opposite the first side wall.
Cover geometry with laterally extending tabs for releasable locking
A cover comprising a main body extending from a first end to a second end, with a first cover tab and a second cover tab extending laterally outward from portions of the first and second ends, wherein releasably locking the cover to the base comprises releasably locking the first cover tab to the first base tab and the second cover tab to the second base tab thereby forming a well around the sample region.
Overall claim coverage focuses on forming a vapor-/fluid-tight well around a sample region by releasably locking corresponding tabs on a cover and base, followed by contacting the sample with a reagent by adding the reagent into the well through an opening formed in the cover. The claim set also includes structural definition of base and cover geometries that support the well formation and reagent delivery.
Stated Advantages
Improve sealing against cross-contamination.
Reduce shear.
Enable uniform heating/condensation control.
Adapt substrates for heating devices.
Improve sealing as described by vapor-/fluid-tight well formation around the sample region.
Documented Applications
Spatial bioassay workflows using a substrate holder with a well around a sample region and reagent contact through a cover opening.
Spatial analysis of analytes in the sample.
Nucleic-acid workflow including permeabilization followed by reverse transcription, second-strand cDNA synthesis, and cDNA library preparation.
Use with glass slide substrates having sample regions with multiple spatially barcoded capture probes.
Tissue processing, staining/immunostaining, in situ hybridization, and RNA-templated ligation (RTL) are described as spatial assay workflow contexts.
Permeabilization agents are described as part of reagent contacting for downstream nucleic-acid workflows.
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