Mechanically induced trapping of molecular interactions
The invention provides devices and methods for surface patterning the substrate of a microfluidic device, and for detection and analysis of interactions between molecules by mechanically trapping a molecular complex while substantially expelling solvent and unbound solute molecules. Examples of molecular complexes include protein-protein complexes and protein-nucleic acid complexes.
1. A method comprising:
in a unit cell of a microfluidic device, said unit cell comprising in a liquid environment:
(1) a microfluidic channel having a substrate,
a molecular complex comprising a first molecule immobilized in a contact area of the substrate, and a second molecule bound to the first molecule and thus indirectly bound to the substrate, and
a movable element that upon actuation contacts the substrate in the contact area wherein the movable element has a geometry such that when it is fully actuated, the contact area extends less than the width of the substrate of the microfluidic channel at the location of the contact area,
actuating the movable element causing it to contact the substrate in the contact area thereby physically trapping the first and second molecules bound to the substrate in the contact area while substantially expelling solvent and solute molecules; or
(2) a microfluidic channel having a substrate,
a first molecule immobilized in a contact area of the substrate,
a second molecule,
a movable element that upon actuation contacts the substrate in the contact area, wherein the movable element has a geometry such that when it is fully actuated, the contact area extends less than the width of the substrate of the microfluidic channel at the location of the contact area,
actuating the movable element causing it to contact the substrate in the contact area thereby physically trapping the first molecule and any second molecules bound to the first molecule substantially expelling solvent and unbound second molecules.
2. The method of claim 1 further comprising de-actuating the movable element.
3. The method of claim 1 wherein the second molecule is labeled with a fluorescent dye.
4. The method of claim 1 further comprising detecting the trapped first and second molecules.
5. The method of claim 4 further comprising detecting a fluorescent signal from said unit cell.
6. The method of claim 1 wherein the movable element is a deflectable elastomeric membrane.
7. The method of claim 6 wherein the membrane is a free-standing membrane that when fully actuated contacts the microfluidic channel substrate without contacting the sides of the microfluidic channel.
8. The method of claim 7 wherein contact between the membrane and the substrate occurs medially and extends radially outward.
9. The method of claim 2 further comprising (i) detecting the first and any second molecules after de-actuation, and (ii) changing the liquid environment surrounding the contact area.
10. The method of claim 9 wherein prior to said de-actuating, the liquid environment in the unit cell is changed.
11. The method of claim 1 wherein the first molecule is an antibody and the second molecule is an antigen.
12. The method of claim 1 wherein the first molecule is a protein and the second molecule is bound by the protein.
13. The method of claim 1 wherein, prior to actuating the movable element, the molecular complex is contacted with a third molecule and the effect of the third molecule on formation or dissociation of the complex is determined.
14. The method of claim 1 carried out on at least 100 unit cells of a microfluidic device, where each of the 100 unit cells comprises a different first molecule.
15. The method of claim 1 carried out on at least 100 unit cells of a microfluidic device, where each of the 100 unit cells comprises a different second molecule.
16. The method of claim 13 carried out on at least 100 unit cells of a microfluidic device, where each of the 100 unit cells comprises a different third molecule.
17. The method of claim 1 , wherein the contact area extends not more than 75% of the width of the microfluidic channel substrate at the location of the contact area when the movable element is fully actuated.
18. The method of claim 17 , wherein the contact area extends not more than 50% of the width of the microfluidic channel substrate at the location of the contact area when the movable element is fully actuated.
19. A method comprising:
in a unit cell of a microfluidic device, said unit cell comprising in a liquid environment:
(1) a microfluidic channel having a substrate,
a molecular complex comprising a first molecule immobilized in a contact area of the substrate, and a second molecule bound to the first molecule and thus indirectly bound to the substrate, and
a movable element that upon actuation contacts the substrate in the contact area of the substrate wherein the movable element when fully actuated does not block flow of fluid through the microfluidic channel,
actuating the movable element causing it to contact the substrate in the contact area thereby physically trapping the first and second molecules bound to the substrate in the contact area while substantially expelling solvent and solute molecules; or
(2) a microfluidic channel having a substrate,
a first molecule immobilized in a contact area of the substrate,
a second molecule,
a movable element that upon actuation contacts the substrate in the contact area of the substrate, wherein the movable element when fully actuated does not block flow of fluid through the microfluidic channel,
actuating the movable element causing it to contact the substrate in the contact area thereby physically trapping the first molecule and any second molecules bound to the first molecule substantially expelling solvent and unbound second molecules.
20. The method of claim 1 , wherein the microfluidic channel is a bulged microfluidic flow channel comprising a widened and rounded portion, and the contact area is located in said widened and rounded portion.
21. The method of claim 6 , wherein the elastomeric membrane has a diameter smaller than the width of the microfluidic channel.
22. A method comprising:
in a unit cell of a microfluidic device, said unit cell comprising in a liquid environment:
(1) a microfluidic channel having a substrate,
a molecular complex comprising a first molecule immobilized in a contact area of the substrate, and a second molecule bound to the first molecule and thus indirectly bound to the substrate, and
a deflectable membrane that upon actuation contacts the substrate in the contact area of the substrate, wherein the contact occurs medially and extends radially outward,
actuating the deflectable membrane causing it to contact the substrate in the contact area thereby physically trapping the first and second molecules bound to the substrate in the contact area while substantially expelling solvent and solute molecules; or
(2) a microfluidic channel having a substrate,
a first molecule immobilized in a contact area of the substrate,
a second molecule,
a deflectable membrane that upon actuation contacts the substrate in the contact area of the substrate, wherein the contact occurs medially and extends radially outward,
actuating the deflectable membrane causing it to contact the substrate in the contact area thereby physically trapping the first molecule and any second molecules bound to the first molecule substantially expelling solvent and unbound second molecules.