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

Isolation Bio Inc

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 are cultured from the at least one cell in the at least one microwell of the array of microwells. The plurality of cells is then analyzed to determine a presence or absence of a biological entity of interest.

Core Innovation

The disclosure describes a microfabricated device having a top surface defining an array of microwells with a surface density of at least 500 microwells per cm2, where at least one microwell includes at least one cell and an amount of a nutrient. A sample is loaded such that the microwells receive cells together with a nutrient amount, and the device supports culturing a plurality of cells from the at least one cell in the at least one microwell. The cultured plurality of cells is then analyzed to determine presence or absence of a biological entity of interest in the sample.

A membrane is applied to retain the at least one cell and the nutrient in the at least one microwell during culturing. The membrane-based retention is positioned as a functional feature of the microwell workflow, including embodiments where the membrane retains cells and the nutrient amount without furnishing additional nutrient during culturing. This microfabricated, membrane-retained microwell array framework is used for culturing and screening biological entities at high microwell density.

The disclosure further provides for selection and downstream handling linked to the microwell identity, including picking at least one cell from a selected microwell of the array and transferring the picked at least one cell to a target location. In related embodiments, analyzing the cultured plurality of cells supports determining whether a biological entity of interest is present or absent, and the microwell selection for picking is driven by that determination. The disclosure also includes embodiments where the analysis and/or correlation of results is associated with microwell location via nucleic-acid-based location tags, including location-specific sequences and target-specific sequences.

Claims Coverage

The partial content includes two independent claims (clm-00001 and clm-00017). Each independent claim centers on a microfabricated device with an array of microwells at a specified high surface density, membrane-retained cell culturing, analysis or selection based on presence/absence of a biological entity of interest, and, depending on the claim, cell picking and transfer to a target location.

Across the two independent claims, the core claim coverage is directed to high-density microfabricated microwell arrays with membrane-retained nutrient/cells for culturing and analyzing to determine presence or absence, and optionally selecting a microwell and transferring picked cells to a target location, with at least one embodiment emphasizing culturing without furnishing additional nutrient.

Stated Advantages

Documented Applications