# US 10,501,716 B2 · Device for mechanical and hydrodynamic microfluidic transfection

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,501,716 B2
- Document ID: US10501716B2
- Status: Granted
- Kind: B2
- Application number: 15/885,766
- Filed: 2018-01-31
- Published: 2018-06-07
- Granted: 2019-12-10
- Earliest claimed: 2015-01-07
- Assignees: Indee. Inc.
- Applicants: Indee. Inc.
- Inventors: Ryan Pawell
- Web page: https://patentgrep.com/patent/US10501716B2

## CPC

- C12M 35/04: {Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli}
- B81B 1/00: Devices without movable or flexible elements, e.g. microcapillary devices
- C12M 23/16: {Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093)}
- C12M 27/18: {Flow directing inserts}
- C12M 35/00: {Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion (machines for extracting juice from animal or plant tissue by electroplasmolysis A23N1/006, processes employing electric or wave energy B01J19/08; treatment of microorganisms or enzymes with electrical or wave energy C12N13/00; methods for cell fusion C12N15/02; introduction of foreign genetic material C12N15/87)}
- C12N 15/87: Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- B81B 2201/058: Microfluidics not provided for in B81B2201/051 - B81B2201/054
- B81B 2203/0338: Channels
- C12N 2510/00: Genetically modified cells
- C12N 2521/00: Culture process characterised by the use of hydrostatic pressure, flow or shear forces
- C12N 2740/15043: viral genome or elements thereof as genetic vector
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- B81B: MICROSTRUCTURAL DEVICES OR SYSTEMS, e.g. MICROMECHANICAL DEVICES (piezoelectric, electrostrictive or magnetostrictive elements per se H10N30/00, H10N35/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Methods for introducing exogenous material into a cell are provided, which include exposing the cell to a transient decrease in pressure in the presence of the exogenous material. Also provided are devices for performing the method of the invention.

## Claims

### Claim 1

A system for introducing exogenous material into cells suspended in a moving fluid through a flow channel of a microfluidic device, comprising:
a substrate;
at least one flow channel within said substrate, said flow channel having at least one inlet, at least one outlet, opposed sidewalls, a width from one of said sidewalls to the other of said sidewalls, and a length perpendicular to the width, said flow channel including a first array of posts along the width of said flow channel, said flow channel including at least a second array of posts along the width of said flow channel; and
a pump configured to move the fluid, cells and exogenous material along the flow channel in a laminar flow to the first array of posts;
wherein the flow channel, arrays of posts, and the pump are configured to cause inertial forces within the fluid to act on the cells in an unsteady flow to permeabilize the cells and introduce the exogenous material into the cells while maintaining at least one viable cell, said second array being shifted a distance relative to said first array to bifurcate the flow coming through said first array, the distance of the shift between said first array and said second array being configured to accommodate the flow of a range of cell sizes, and the flow around each post in a region of the flow channel has an object Reynolds number of at least 40.

### Claim 2 (depends on 1)

The system of claim 1, wherein each post has a cylindrical cross section.

### Claim 3 (depends on 1)

The system of claim 1, wherein said second array of posts is parallel to said first array of posts.

### Claim 4 (depends on 1)

The system of claim 1, wherein a plurality of said posts are completely spaced apart from said opposed walls of said flow channel.

### Claim 5 (depends on 1)

The system of claim 1, wherein each post has a height greater than its maximum width.

### Claim 6 (depends on 1)

The system of claim 1, wherein each post has a height equal to its maximum width.

### Claim 7 (depends on 1)

The system of claim 1, wherein the substrate is a fused silica.

### Claim 8 (depends on 1)

The system of claim 1, wherein the substrate is an ion-etched fused silica bonded to a laser-machined fused silica wafer.