Nanopore Replacement via Osmotic Imbalance in Sequencing Membranes
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Solution Overview
Problem
Existing methods for removing and replacing nanopores in nanopore-based sequencing chips are not compatible with the hydrostatic and electromotive forces present in chip-based sensor arrays, leading to membrane destruction and increased complexity in the sequencing process.
Innovation Solution
A technique involving the use of electrolyte solutions with different osmolarities to nondestructively eject and replace nanopores in a nanopore-based sequencing chip. This is achieved by flowing a first electrolyte solution with a different osmolarity than the initial osmolarity of the reservoir, causing the membrane to bow and eject the nanopore, followed by the introduction of a second electrolyte solution with replacement nanopores and a similar osmolarity to the initial reservoir, allowing for the insertion of new nanopores.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If the patch-clamp method is used to remove and replace nanopores, then nanopore replacement is enabled, but the hydrostatic and electromotive forces are not compatible with chip-based sensor arrays causing membrane destruction
Solution Approach 1:
The patent changes the physical parameters of the electrolyte solution, specifically the osmolarity, to enable nanopore ejection. By introducing an osmotic gradient through electrolyte solutions of different osmolarities, the membrane experiences osmotic pressure that causes it to bow and eject the nanopore without requiring the destructive forces of the patch-clamp method.
Solution Approach 2:
The patent replaces the mechanical and electromotive forces of the patch-clamp method with an osmotic-based mechanism. Instead of using hydrostatic pressure and electrical fields that destroy the membrane, the invention uses osmotic pressure generated by electrolyte solution concentration differences to achieve nanopore removal and replacement.
2Ease of operation
If standard techniques are used to remove nanopores from lipid bilayer membranes, then nanopore removal is achieved, but the bilayer must be broken and reformed increasing complexity
Solution Approach 1:
The patent extracts the nanopore from the membrane using osmotic pressure without destroying the membrane itself. The electrolyte solution with different osmolarity creates an osmotic gradient that causes the membrane to bow and eject the nanopore, allowing the nanopore to be removed while the membrane remains intact and functional.
Solution Approach 2:
The patent introduces an intermediary substance (electrolyte solution with specific osmolarity) that mediates the nanopore removal process. This intermediary creates the osmotic gradient necessary for nanopore ejection without requiring direct mechanical or chemical interaction that would damage the membrane.
3Productivity
If nanopores are removed and replaced in sequencing chips, then additional nucleic acid sequencing is enabled, but existing methods cause membrane destruction
Solution Approach 1:
The patent changes the osmolarity parameter of the electrolyte solution to enable repeated nanopore removal and replacement cycles. By controlling the osmotic gradient through electrolyte concentration, the system can eject and replace nanopores multiple times without degrading the membrane, thereby increasing sequencing throughput while maintaining membrane stability.
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 enables the nondestructive removal and replacement of nanopores, increasing the throughput of single molecule sensor arrays and nanopore-based sequencing chips by avoiding membrane destruction and simplifying the sequencing process.
Implementation Method 1
The flowing of the first electrolyte solution to the first reservoir changes the first electrolyte reservoir osmolarity from a first initial osmolarity to a new osmolarity that is different from the second initial osmolarity. The difference between the new and second initial osmolarities causes the membrane to bow in the direction of the second electrolyte reservoir, and the nanopore to eject from the membrane as a result of this bowing.
Implementation Method 2
One of the plurality of replacement nanopores is then inserted into the membrane. In one embodiment, the insertion comprises applying an electroporation voltage across the membrane.
Data Source
AI summary
Techniques for replacing nanopores within a nanopore based sequencing chip are provided. A first electrolyte solution is added to the external reservoir of the sequencing chip, introducing an osmotic imbalance between the reservoir and the well chamber located on the opposite side of a lipid bilayer membrane. The osmotic imbalance causes the membrane to change shape, and a nanopore within the membrane to be ejected. A second electrolyte solution is then added to the external reservoir to provide replacement nanopores and to restore the membrane shape. The replacement nanopores can be inserted into the membrane, effectively replacing the initial pore without causing the destruction of the membrane.