High resolution systems, kits, apparatus, and methods for high throughput microbiology applications
Inventors
Blainey, Paul C. • Seely, Michael W. • Stocker, Roman • Zengler, Karsten • Conradson, Scott • Christey, Peter • Hallock, Alexander
Assignees
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Publication Number
US-11104940-B2
Publication Date
2021-08-31
Expiration Date
Abstract
A method of screening for at least one biological entity of interest using a microfabricated device which has a top surface defining an array of microwells. A sample is loaded onto the microfabricated device such that at least one microwell of the array of microwells includes at least one cell and an amount of a nutrient. A membrane is applied to the microfabricated device to retain the at least one cell and the nutrient. A plurality of cells is cultured from the at least one cell in the at least one microwell of the array of microwells. The plurality of cells in the at least one microwell of the array of microwells are split into a first portion of the plurality of cells and a second portion of the plurality of cells. The plurality of cells is analyzed to determine a presence or absence of a biological entity of interest.
Core Innovation
The invention relates to culturing and screening for at least one biological entity using a microfabricated device with a top surface defining an array of microwells. A sample is loaded so that at least one microwell includes at least one cell and an amount of a nutrient, and a membrane is applied to retain the at least one cell and the nutrient in the microwell while cells are cultured from the at least one cell.
After culturing a plurality of cells from the microwell, the plurality of cells in the microwell is split into a first portion and a second portion. The method analyzes the plurality of cells to determine a presence or absence of a biological entity of interest in the sample, enabling screening tied to the microwell’s cultured lineage and its split portions.
In another aspect, the approach includes picking at least one cell from a selected microwell of the array and transferring the picked cell to a target location. The selected microwell is associated with the microwell that is selected based on downstream determinations, while the overall workflow maintains spatial separation via the microwell array and retains nutrient and cells using the membrane.
Claims Coverage
The document provides two independent method claims. Across the independent claims, the inventive features center on high-density microfabricated microwell arrays, membrane-based retention of cells and nutrient, splitting of cultured cell populations into portions, and either analyzing for presence or absence or picking from a selected microwell for transfer to a target location.
Microfabricated microwell array for culturing with membrane retention
A method using a microfabricated device having a top surface defining an array of microwells, loading a sample such that at least one microwell includes at least one cell and an amount of a nutrient, applying a membrane to retain the at least one cell and the nutrient in the microwell, and culturing a plurality of cells from the at least one cell in the microwell array.
Splitting cultured cell plurality into first and second portions for screening
Splitting the plurality of cells in the at least one microwell of the array of microwells into a first portion of the plurality of cells and a second portion of the plurality of cells.
Analyzing for presence or absence of biological entity of interest
Analyzing the plurality of cells to determine a presence or absence of a biological entity of interest in the sample.
Picking from a selected microwell and transferring to a target location
Picking at least one cell from the plurality of cells from a selected microwell of the array of microwells and transferring the picked at least one cell to a target location.
Across the independent claims, the inventive coverage is anchored in a microfabricated microwell array combined with membrane retention of cells and nutrient, followed by culturing and splitting of the cultured plurality of cells. Screening is performed either by analyzing the plurality of cells for presence or absence of a biological entity of interest, or by selecting a microwell for cell picking and transfer of picked cells to a target location.
Stated Advantages
Documented Applications
Biobanking cells, including culturing and splitting into portions for downstream identification and screening workflows.
Microbiome and soil screening using presence/absence screening workflows tied to microwell culturing.
Cultivation and adaptation of microorganisms using microfabricated high-density microwell culturing with membrane-based nutrient and cell retention.
Droplet-based alternatives are mentioned as an alternative approach.
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