Osmotic imbalance methods for bilayer formation
A method of forming a plurality of lipid bilayers over an array of cells in a nanopore based sequencing chip is disclosed. Each of the cells comprises a well. A first salt buffer solution with a first osmolarity is flowed over a cell in the nanopore based sequencing chip to substantially fill a well in the cell with the first salt buffer solution. A lipid and solvent mixture is flowed over the cell to deposit a lipid membrane over the well that encloses the first salt buffer solution in the well. A second salt buffer solution with a second osmolarity is flowed above the well to reduce the thickness of the lipid membrane, wherein the second osmolarity is a lower osmolarity than the first osmolarity such that an osmotic imbalance is created between a first volume inside the well and a second volume outside the well.
1. A method of forming a plurality of lipid bilayers over an array of cells in a nanopore based sequencing chip, each of the cells comprising a well, the method comprising:
flowing a first salt buffer solution with a first osmolarity over a cell in the nanopore based sequencing chip to substantially fill a well in the cell with the first salt buffer solution;
flowing a lipid and solvent mixture over the cell to deposit a lipid membrane over the well that encloses the first salt buffer solution in the well; and
flowing a second salt buffer solution with a second osmolarity above the well to reduce the thickness of the lipid membrane to form a lipid bilayer, wherein the second osmolarity is a lower osmolarity than the first osmolarity such that an osmotic imbalance is created between a first volume inside the well and a second volume outside the well;
wherein the step of flowing the second salt buffer solution with the second osmolarity above the well is performed in a number of cycles over time, and wherein the second osmolarity is progressively increased in the number of cycles.
2. The method of claim 1 , wherein the osmotic imbalance causes water to diffuse through the lipid membrane into the well and causes the lipid membrane to bow upwards.
3. The method of claim 1 , further comprising applying a lipid bilayer initiating stimulus to facilitate creation of a small lipid bilayer on the lipid membrane.
4. The method of claim 3 , wherein the step of applying the lipid bilayer initiating stimulus is performed in a number of cycles over time, and wherein the lipid bilayer initiating stimulus level is adaptable in the number of cycles.
5. The method of claim 3 , wherein the lipid bilayer initiating stimulus comprises a vibration stimulus.
6. The method of claim 5 , wherein applying the vibration stimulus comprises generating waves in the second volume outside the well.
7. The method of claim 3 , wherein the lipid bilayer initiating stimulus comprises an electrical stimulus.
8. An apparatus for forming a plurality of lipid bilayers over an array of cells in a nanopore based sequencing chip, the system comprising:
a nanopore based sequencing chip comprising an array of cells, each of the cells comprising a well;
a flow chamber coupled to the nanopore based sequencing chip; and
a processor and memory for storing instructions that, when executed by the processor, is configured to:
flow a first salt buffer solution with a first osmolarity over a cell in the nanopore based sequencing chip to substantially fill a well in the cell with the first salt buffer solution;
flow a lipid and solvent mixture over the cell to deposit a lipid membrane over the well that encloses the first salt buffer solution in the well; and
flow a second salt buffer solution with a second osmolarity above the well to reduce the thickness of the lipid membrane to form a lipid bilayer, wherein the second osmolarity is a lower osmolarity than the first osmolarity such that an osmotic imbalance is created between a first volume inside the well and a second volume outside the well;
wherein the step of flowing the second salt buffer solution with the second osmolarity above the well is performed in a number of cycles over time, and wherein the second osmolarity is progressively increased in the number of cycles.
9. The apparatus of claim 8 , wherein the osmotic imbalance causes water to diffuse through the lipid membrane into the well and causes the lipid membrane to bow upwards.
10. The apparatus of claim 8 , wherein the processor or the circuitry is further configured to apply a lipid bilayer initiating stimulus to facilitate creation of a small lipid bilayer on the lipid membrane.
11. The apparatus of claim 10 , wherein the step of applying the lipid bilayer initiating stimulus is performed in a number of cycles over time, and wherein the lipid bilayer initiating stimulus level is adaptable in the number of cycles.
12. The apparatus of claim 10 , wherein the lipid bilayer initiating stimulus comprises a vibration stimulus.
13. The apparatus of claim 10 , wherein the lipid bilayer initiating stimulus comprises an electrical stimulus.