Microfluidic Channel Fluidic Pump Cell Transfection
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Solution Overview
Problem
Current cell transfection methods, such as viral transfection and lipofection, are laborious and introduce unwanted contaminants or size limitations, while electrotransfection is inefficient for exploring transfection conditions at the single cell level and for large molecules like CRISPR-Cas9 or nanoparticles.
Innovation Solution
A mechanical transfection method using a fluidic pump within a microfluidic channel creates shear forces to form apertures in cell membranes, allowing for efficient introduction of transfection materials like nucleic acids, proteins, and nanoparticles without contaminants, enabling single-cell level processing and optimization of transfection conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If viral transfection or lipofection is used, then transfection material can be introduced into cells, but the process is laborious and introduces unwanted contaminants
Solution Approach 1:
The patent replaces viral or chemical transfection methods with a mechanical system consisting of a microfluidic channel and fluidic pump that applies controlled shear force to cells. This mechanical approach introduces no biological contaminants (unlike viral vectors) and no chemical surfactants (unlike lipofection), while maintaining effective transfection through physically-formed apertures in the cell membrane.
Solution Approach 2:
The patent extracts and eliminates the harmful components (viral vectors, chemical surfactants) from the transfection process, retaining only the essential mechanical action of forcing material through the cell membrane via controlled shear force in a clean microfluidic environment.
2Adaptability or versatility
If electrotransfection is used, then transfection can be performed, but it is inefficient for single cell level processing and large molecules
Solution Approach 1:
The patent segments the transfection process to the single-cell level by flowing individual cells through a microfluidic channel one at a time, where each cell experiences controlled shear force and forms apertures for material uptake. This enables precise, cell-by-cell transfection that is highly efficient for both single-cell applications and large macromolecules like CRISPR-Cas9 and nanoparticles.
Solution Approach 2:
The patent changes the transfection mechanism from electrical fields (electrotransfection) to controlled mechanical shear force in a microfluidic environment. This parameter change enables better handling of large molecules and single-cell processing, as the mechanical aperture formation is size-independent and can be precisely controlled at the single-cell level.
3Ease of operation
If mechanical transfection with fluidic pump is used, then single cell level processing is enabled, but device complexity increases
Solution Approach 1:
The patent uses a fluidic pump to generate controlled fluid flow through a microfluidic channel, creating shear force on cells as they pass through a constriction region. This hydraulic approach enables automated single-cell transfection with precise control over flow rate and shear force, making the system easier to operate despite the microfluidic complexity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method allows for precise and efficient transfection of cells at the single-cell level, optimizing transfection conditions and accommodating various types of cells and materials, without introducing contaminants, and enables parallel processing and assessment of transfection efficiency.
Implementation Method 1
A fluidic pump of the apparatus may be actuated to cause a cell of the biologic sample to flow into and through a constriction region of the microfluidic channel... The flow of the cell through the constriction region may cause the cell to form apertures in the cell membrane
Data Source
AI summary
An example apparatus comprises includes a first reservoir to store a biologic sample containing a cell, a microfluidic channel fluidically coupled to the first reservoir, and circuitry. The microfluidic channel includes a constriction region including a first circumference that is attenuated from remaining portions of the microfluidic channel, and a fluidic pump disposed within the microfluidic channel. The circuitry is to activate the fluidic pump to direct flow of the cell from the first reservoir to the microfluidic channel and through the constriction region.