Method for mechanical and hydrodynamic microfluidic transfection and apparatus therefor
Methods for introducing exogenous material into a cell are provided, which include exposing the cell to a transient decrease in pressure in the presence of the exogenous material. Also provided are devices for performing the method of the invention.
1. A method for introducing an exogenous material into a cell, comprising:
introducing a liquid including the cell and the exogenous material into a flow channel of a microfluidic device, the channel including at least two flow diverters, the gap between the at least two flow diverters having a width, the width of the gap being greater than the diameter of the cell; and
exposing said cell to a transient decrease in pressure and unsteady flow downstream of the flow diverters when the cell flows past the flow diverters to thereby introduce said exogenous material into said cell.
2. The method of claim 1 , wherein the cell is viable after being exposed to the transient decrease in pressure and unsteady flow.
3. The method of claim 1 , wherein said exogenous material is selected from the group consisting of a small organic molecule, a nucleic acid, a nucleotide, an oligonucleotide, a protein, a peptide, an amino acid, a lipid, a polysaccharide, a quantum dot, a carbon nanotube, a nanoparticle, a gold particle, a monosaccharide, a vitamin and a steroid.
4. The method of claim 1 , wherein said exogenous material is introduced into the cytoplasm of said cell.
5. The method of claim 1 , wherein said cell is a mammalian cell, a yeast cell or an insect cell and said exogenous material is introduced into the nucleus of said cell.
6. The method of claim 1 , wherein said cell is exposed to said transient decrease in pressure in the presence of said exogenous material for at least 10 nanoseconds.
7. The method of claim 1 , wherein unsteady flow is at least one of a laminar vortex street, a transitional vortex street, a turbulent vortex street, transitional flow or turbulent flow.
8. A method for introducing an exogenous material into a cell, comprising:
introducing a liquid including the cell and the exogenous material into a flow channel of a microfluidic device, the channel including at least two flow diverters, the gap between the at least two flow diverters having a width, the width of the gap being greater than the diameter of the cell; and
exposing said cell to a transient decrease in pressure and unsteady flow downstream of the flow diverters when the cell flows past the flow diverters to thereby introduce said exogenous material into said cell, wherein said transient decrease in pressure is at least 10 kPa.
9. The method of claim 1 , wherein each flow diverter is a post.
10. The method of claim 9 , wherein the post is cylindrical.
11. The method of claim 10 , wherein the post has a diameter greater than the gap between the posts.
12. The method of claim 10 , wherein the post has a diameter equal to the gap between the posts.
13. The method of claim 10 , wherein the post has a diameter less than the gap between the posts.
14. The method of claim 1 , wherein each flow diverter has a maximum width of 10 micrometers.
15. The method of claim 1 , wherein each flow diverter has a maximum width of 100 micrometers.
16. The method of claim 1 , wherein each flow diverter has a maximum width of 1 millimeter.
17. A method for transfecting a cell to introduce an exogenous material into the cell, comprising:
introducing a liquid including the cell and the exogenous material into a flow channel of a microfluidic device, the flow channel having a width and at least two flow diverters oriented in an array along the width of the flow channel, each flow diverter having a maximum width, the maximum width of each flow diverter being greater than the gap between the at least two flow diverters; and
exposing the cell to a transient decrease in pressure and unsteady flow downstream of the flow diverters when the cell flows past the flow diverters to thereby introduce the exogenous material into the cell.
18. The method of claim 17 , wherein the at least two flow diverters include a plurality of posts.
19. The method of claim 17 , wherein the maximum width of each flow diverter is 10 micrometers.
20. The method of claim 17 , wherein each flow diverter has a height greater than its maximum width.
21. The method of claim 1 , wherein the exogenous material introduced into the cell is an expression vector.
22. The method of claim 21 , wherein the expression vector is a bacterial artificial chromosome (BAC).
23. The method of claim 1 , wherein the cell is a mammalian cell having a nucleus and the exogenous material is introduced into the nucleus.
24. The method of claim 1 , wherein the cell is exposed to the transient decrease in pressure and unsteady flow in the presence of the exogenous material for at least 10 nanoseconds.
25. The method of claim 1 , wherein the flow has a maximum peak velocity of at least 1 meter per second.
26. The method of claim 1 , wherein the flow of the liquid around the flow diverters has an object Reynolds number of at least 40.
27. The method of claim 17 , wherein the cell is exposed to the transient decrease in pressure and unsteady flow in the presence of the exogenous material for at least 10 nanoseconds.
28. The method of claim 17 , wherein the flow has a maximum peak velocity of at least 1 meter per second.
29. The method of claim 17 , wherein the flow of the liquid around the flow diverters has an object Reynolds number of at least 40.
30. The method of claim 8 , wherein the flow of the liquid around the flow diverters has an object Reynolds number of at least 40.