Flow Electroporation Sheath Flow Layout for High-Concentration Suspensions
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Solution Overview
Problem
Existing flow electroporation methods face efficiency issues when the concentration of biologically derived substances is high, leading to reduced introduction and utilization of bioactive substances.
Innovation Solution
The method involves forming sheath flows with a sheath liquid and setting the thickness of the suspension flow between electrodes to 1 to 10 mm, combining it with sheath flows downstream, and adjusting the electric field and pulse width based on conductivity and concentration measurements, while maintaining a dilution rate of 2 times or less.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If the concentration of biologically derived substance in the suspension is increased to improve utilization efficiency of bioactive substance, then the introduction efficiency decreases due to retention near channel walls
Solution Approach 1:
A sheath liquid is introduced as an intermediary substance that flows between the suspension and the electrode surfaces. This sheath liquid prevents direct contact between high-concentration biologically derived substances and the channel walls, eliminating retention while maintaining high suspension concentration for improved utilization efficiency.
Solution Approach 2:
The invention uses hydraulic flow control by adjusting the flow rates of both the suspension and sheath liquid to optimize the flow dynamics. By controlling the hydraulic parameters, the system maintains laminar flow patterns that prevent retention of biologically derived substances near the channel walls while ensuring efficient electroporation.
2Manufacturing precision
If the thickness of suspension flow is reduced to improve electric field uniformity, then the throughput decreases
Solution Approach 1:
The sheath liquid acts as a mediator that enables the use of thicker suspension flow layers. By preventing direct contact between the suspension and electrode surfaces, the sheath liquid allows increased suspension flow thickness while maintaining electric field uniformity, thereby improving throughput without sacrificing precision.
Solution Approach 2:
The invention changes the physical parameters of the system by introducing a sheath liquid with different electrical properties. This parameter change allows the suspension flow thickness to be increased while maintaining the electric field uniformity required for effective electroporation, resolving the contradiction between precision and productivity.
3Productivity
If sheath liquid flow rate is increased to prevent retention of biologically derived substances, then the dilution rate increases
Solution Approach 1:
The invention optimizes the flow rate parameters of the sheath liquid to achieve the minimum necessary flow to prevent retention of biologically derived substances. By carefully controlling this parameter, the system maintains high introduction efficiency while minimizing dilution of the suspension, thus resolving the contradiction between productivity and substance concentration.
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 approach enables high-efficiency electroporation with improved throughput, introduction efficiency, and utilization of bioactive substances, even at high concentrations, by preventing retention of biologically derived substances near the channel walls and ensuring uniform electric field application.
Implementation Method 1
An electric perforating method is a method of introducing a substance into cells by perforating holes in a cell membrane with an electric pulse
Implementation Method 2
forming sheath flows of a sheath liquid which flows together with the suspension and comes into contact with electrodes constituting the electrode pair
Implementation Method 3
the bioactive substance passes through the membrane with increased permeability by diffusion or electrophoresis
Implementation Method 4
the bioactive substance passes through the membrane with increased permeability by diffusion or electrophoresis
Data Source
AI summary
An object is to provide an electroporation method capable of performing electroporation with high efficiency, a method for manufacturing a useful substance using the method, a flow channel device for performing the method, and an electroporation apparatus using the flow channel device. The object is achieved by, in the electroporation, forming sheath flows which flow together with a suspension and comes into contact with an electrode, and further having a restriction region which restricts a thickness of a suspension flow consisting of the suspension in a separation direction of an electrode pair to 1 to 10 mm.