Systems And Methods For Transfer Of Reagents Between Droplets
Patent No. US12416102 (titled "Systems And Methods For Transfer Of Reagents Between Droplets") was filed by 10X Genomics Inc on Aug 9, 2021.
What is this patent about?
’102 is related to the field of droplet microfluidics and single-cell analysis. Specifically, it addresses the challenge of controlling reagent concentrations within droplets used for processing biological samples, such as cells, for downstream analysis like sequencing. The background involves partitioning biological samples into droplets to process them separately, enabling accurate control of the reaction environment. However, changing the amount of reagent in a droplet, such as increasing or decreasing the concentration, remains challenging.
The underlying idea behind ’102 is to use a mediator to transfer a reagent from one droplet to another. This mediator, comprising an aggregate of surfactant molecules, facilitates the transport of the reagent between droplets, allowing for controlled changes in reagent concentration within the droplets. This is particularly useful when the reagent is needed to trigger a specific event within the droplet, such as releasing nucleic acid molecules from an analyte carrier.
The claims of ’102 focus on a method for nucleic acid processing that involves generating a plurality of droplets by agitating a first fluid volume containing beads and analyte carriers with a second immiscible fluid volume. The key aspect is subjecting a droplet containing a bead and an analyte carrier to conditions that allow a reagent to be transferred to it from another droplet via a surfactant aggregate mediator . This reagent is specifically configured to release nucleic acid molecules from the analyte carrier, enabling the generation of barcoded nucleic acid molecules.
In practice, the invention involves creating two populations of droplets: one containing the analyte carriers and beads, and another containing the reagent. These populations are then mixed, and the reagent is transferred from the second droplet population to the first via the surfactant aggregate mediator. This mediator could be a micelle formed by surfactant molecules in the continuous phase surrounding the droplets. The reagent, once transferred, acts on the analyte carrier to release nucleic acid molecules, which are then barcoded using the nucleic acid barcode molecules on the bead.
This approach differs from prior methods by providing a mechanism for dynamically adjusting reagent concentrations within droplets after droplet formation. Instead of relying on precise initial reagent loading, the invention allows for post-formation reagent delivery , offering greater flexibility and control over the reaction environment. This is particularly advantageous for reactions that require specific reagent concentrations or timed reagent delivery, such as those involved in single-cell sequencing workflows.
How does this patent fit in bigger picture?
Technical landscape at the time
In the early 2020s when ’102 was filed, single-cell analysis at a time when single-cell analysis was typically implemented using microfluidic devices to partition cells into droplets, where each droplet ideally contained a single cell and a set of reagents for downstream processing such as barcoding and sequencing. When systems commonly relied on precise control over reagent concentrations within these droplets to ensure efficient and accurate reactions, and when hardware or software constraints made controlling reagent concentrations within droplets non-trivial.
Novelty and Inventive Step
The examiner approved the application because it was found that the prior art, specifically Thiam et al., does not explicitly teach a method for nucleic acid processing that includes generating droplets by agitating a first fluid volume containing beads with barcode molecules and analyte carriers with nucleic acid molecules, mixing this with a second immiscible fluid, and then transferring a reagent between droplets via a mediator to release nucleic acid molecules for barcoding.
Claims
This patent contains 21 claims, with claim 1 being the only independent claim. Independent claim 1 is directed to a method for nucleic acid processing using droplets, beads with barcode molecules, and analyte carriers. The dependent claims generally elaborate on specific components, conditions, or steps within the method outlined in the independent claim.
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Date
Description
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US12416102
- Application Number
- US17397775
- Filing Date
- Aug 9, 2021
- Status
- Granted
- Expiry Date
- Dec 11, 2042
- External Links
- Slate, USPTO, Google Patents