Georgia Tech Research Corporation

Cellfe, Inc.

Alexander Alexeev

Todd A Sulchek

Sewoon Han

Gonghao Wang

Ian Sicher

Bushra Tasadduq

Ockchul Kim

Miguel Calero-garcia

Alla Zamarayeva

Anna Liu

The present disclosure provides methods and systems for cell processing, including delivery of imaging agents into cells. The methods and systems may comprise the use of a microfluidic device. The microfluidic device may comprise a channel comprising a compressive element. The compressive element may be configured to reduce a volume of the cell and facilitate the formation of one or more transient pores in a cell membrane of the cell. The one or more pores may permit one or more imaging agents to enter the cell. Also provided are modified cells produced using the disclosed methods and systems and methods of imaging the modified cells in a subject.

Methods and systems for cell labeling and imaging

Described herein are methods and systems for mechanoporation-based high-throughput payload delivery into biological cells. For example, one system can process at least 1 billion cells per minute or at least 25 billion cells per minute, which is substantially greater than conventional methods. A cell processing apparatus comprises a processing assembly formed by stacking multiple processing components. Each processing component comprises channels, which may be used for filtration, mechanoporation, and/or separation of cells in the cell media. This functionality depends on the configuration of each channel. For example, each channel comprises one or more ridges such that each ridge forms a processing gap with an adjacent one of the processing components. The ridges may extend to the side walls or form a bypass gap with the wall. The processing gaps can be specially configured to compress cells as the cells pass through these gaps thereby initiating the mechanoporation process.

Methods and systems for mechanoporation-based high-throughput payload delivery into biological cells

Described herein are methods and systems for cell processing or, more specifically, for introducing various payloads into cells. These methods and systems use a mechanoporation approach in which cells are rapidly compressed and then released to relax while absorbing the payload. More specifically, these methods and systems enable high-throughput mechanoporation with various clogging mitigation features. A cell processing apparatus comprises a shell with an inner shell cylindrical surface, a core with an outer core cylindrical surface, and ridges, supported on and protruding away from one of the inner shell cylindrical surface and the outer core cylindrical surface. The core is disposed inside the shell. The outer core cylindrical surface is concentric with the inner shell cylindrical surface. Each of the ridges forms a ridge gap with the other one of the inner shell cylindrical surface and the outer core cylindrical surface.

Methods and systems for high-throughput cell processing

Microfluidic devices for cell sorting or cell fractionation are disclosed. A microfluidic device can comprise one or more inlets, a first wall and a second wall, and two or more outlets. The first and second walls can be substantially planar to each other and the first wall having can have a plurality of ridges protruding from the first wall and defining a compression gap between the ridge and a surface of the second wall. The microfluidic device can also be a cell sorting device for sorting a plurality of cells based on one or more biophysical cellular properties including size, elasticity, viscosity, and/or viscoelasticity wherein the cells are subjected to one or more compressions due to the compression gap. Also disclosed are methods for cell sorting based on a variety of biophysical cellular properties.

Microfluidic devices for cellular sorting

A microchannel for processing cells by compression of the cells including an inlet, ridges and an outlet. Each ridge including a compressive surface and a cell adhesion entity. The outlet configured to remove at least one of a first portion of the cells and a second portion of the cells from the microchannel. Each ridge oriented at an angle of from 25 degrees to 70 degrees relative to a center axis of the microchannel. The cell adhesion entity configured such that the first portion of the cells has a first adhesion property relative to the cell adhesion entity to follow a first trajectory through the microchannel. The cell adhesion entity further configured such that the second portion of the cells has a second adhesion property relative to the cell adhesion entity to follow a second trajectory through the microchannel. The first trajectory is different from the second trajectory.

Systems and methods for continuous sorting of cells based on molecular adhesion

Embodiments of the present disclosure can include a method comprising: providing a plurality of cells to a microchannel, the microchannel coated in at least one cell adhesion entity and comprising a compressive surface and a first outlet, the compressive surface defining a compression gap, flowing the plurality of cells through the microchannel, wherein the flowing comprises: compressing the plurality of cells underneath the compressive surface; and exposing the plurality of cells to the at least one cell adhesion entity, wherein the exposing causes a first portion of the cells having a first adhesion property to temporarily bind to the cell adhesion entity; and collecting the first portion of cells at the first outlet; wherein the compression gap has a height of from 75% to 95% an average diameter of the plurality of cells.

Methods for continuous sorting of cells based on molecular adhesion

Embodiments of the present disclosure can include a method for convective intracellular delivery including providing cells and molecules to a microchannel having compressive surfaces, wherein the compressive surfaces define compression gaps having a height of from 20 and 80% of the average cell diameter; and a plurality of relaxation spaces disposed between the compressive surfaces; flowing the cell medium through the microchannel, wherein as the cell medium flows through the microchannel, the plurality of cells undergo a convective intracellular delivery process comprising: compressing the plurality of cells, wherein the compressing causes the plurality of cells to undergo a loss in intracellular volume (V); and passing the plurality of cells to a first relaxation space, wherein the plurality of cells undergo a gain in volume (V) and absorb a portion of the plurality of molecules.

Methods for convectively-driven intracellular delivery

An apparatus for separating particles includes a first planar wall and a spaced apart second planar wall parallel to the first planar wall. The first planar wall and the second planar wall define a passage therebetween, which is disposed along a fluid flow axis. A first plurality of spaced apart elongated ridges extends into the passage from the first planar wall. The first plurality of spaced apart elongated ridges is disposed along a diagonal direction relative to the fluid flow axis. When a fluid is moved through the passage in a direction corresponding to the fluid flow axis, particles of a first type will tend to move in a first direction that is diagonally away from the fluid flow axis and particles of a second type, different from the first type, will tend to move in a second direction that is different from the first direction.

Growing community of inventors

Atlanta, GA, United States of America

Alexander Alexeev