PLANAR LIPID BILAYER ARRAY FORMED BY MICROFLUIDIC TECHNIQUE AND METHOD OF ANALYSIS USING PLANAR LIPID BILAYER

• US 20100304980A1
• Filed: 11/26/2008
• Published: 12/02/2010
• Est. Priority Date: 11/26/2007
• Status: Active Grant

• Current Assignee: University of Tokyo
• Original Assignee: University of Tokyo
• Inventors: Tan, Wei-Heong, Ota, Sadao, Suzuki, Hiroaki, Takeuchi, Shoji
• Granted Patent: US 8,513,165 B2
• Time in Patent Office: Days
• US Class Current: 506/7
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/041 Connecting closures to device or container
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: C40B 60/00 Apparatus specially adapted for use in combinatorial chemistry or with libraries
• CPC Class Codes: G01N 2405/04 Phospholipids, i.e. phosphoglycerides
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

There is provided a planar lipid bilayer array formed by microfluidic technique and a method of analysis using the planar lipid bilayers, providing the advantages such as portability, decreased analysis time, a smaller amount of required reagents, and parallel automation with high reproducibility. The planar lipid bilayer array formed by microfluidic technique is a planar lipid bilayer array formed by microfluidic technique (PDMS device) 1 saturated with water by preliminarily immersing in water, comprising microchannels 2 connected to an inlet of a microfluidic channel and arranged in parallel, and microchambers 3 having apertures on both sides of the microchannel 2.

Claims

1. A planar lipid bilayer array formed by microfluidic technique, wherein the planar lipid bilayer array comprises:

(i) microchannels connected to an inlet of a microfluidic channel and arranged in parallel; and

(ii) microchambers having apertures on both sides of the microchannel.

2. The planar lipid bilayer array formed by microfluidic technique according to claim 1, wherein the apertures on the both sides of the microchannel are arranged in a zigzag manner in relation to the microchannel.

3. The planar lipid bilayer array formed by microfluidic technique according to claim 1, wherein the planar lipid bilayer array is an array saturated with water by preliminarily immersing in water.

4. The planar lipid bilayer array formed by microfluidic technique according to claim 3, wherein the array is configured by PDMS.

5. The planar lipid bilayer array formed by microfluidic technique according to claim 1, wherein the both sides of the microchannel are both sidewalls of the microchannel.

6. The planar lipid bilayer array formed by microfluidic technique according to claim 1, wherein the both sides of the microchannel are top and bottom walls of the microchannel.

7. The planar lipid bilayer array formed by microfluidic technique according to claim 1, wherein the microchannel is 7 μ

m in height, and the microchamber is 17 μ

m in width and 19 μ

m in height.

8. The planar lipid bilayer array formed by microfluidic technique according to claim 1, comprising a single microsyringe and a single tube connected to the microsyringe, wherein a buffer solution containing membrane protein and fluorescent material, an organic solvent containing lipid, and a buffer solution containing no dissolved materials are infused sequentially into the tube from the tip, and the tip of the tube is connected to the inlet of the microfluidic channel.

9. A method of analysis using planar lipid bilayers, comprising the steps of:

(a) preliminarily immersing a planar lipid bilayer array formed by microfluidic technique in water to saturate with water; and

(b) with the use of microchannels arranged in parallel in the planar lipid bilayer array saturated with water and microchambers having apertures on both sides of the microchannel, forming the planar lipid bilayers in the apertures to seal the microchambers.

10. The method of analysis using planar lipid bilayers according to claim 9, wherein the apertures on the both sides of the microchannels are arranged in a zigzag manner in relation to the microchannels.

11. The method of analysis using planar lipid bilayers according to claim 9, comprising the steps of:

(a) infusing a buffer solution containing membrane protein and fluorescent material into the microchannels and the microchambers;

(b) infusing an organic solvent containing lipid into the microchannels; and

(c) infusing a buffer solution containing no dissolved materials into the microchannels.

12. The method of analysis using planar lipid bilayers according to claim 11, wherein the membrane protein is α

-Hemolysin, the fluorescent material is Calcein, the lipid is phosphatidycoline (PC), and the organic solvent is hexadecane.

13. The method of analysis using planar lipid bilayers according to claim 11, wherein the membrane protein is α

-Hemolysin, the fluorescent material is Calcein, the lipid is phosphatidycoline (PC), and the organic solvent is squalene.

14. A method of analysis using planar lipid bilayers, wherein a degree of membrane transport of the planar lipid bilayer is measured by deforming the planar lipid bilayer fabricated according to claim 11, forming a liposome, and introducing Hemolysin into the liposome.

15. The method of analysis using the planar lipid bilayers according to claim 14, wherein a degree of decrease in the fluorescence intensity is measured in the liposome containing fluorescent material in addition to Hemolysin.