Sonication to selectively lyse different cell types
View Patent ↗A sonication apparatus is directed to a microfluidic-based system to automate differential extraction of specific cell types within a mixed sample. The microfluidic-based system includes a sonication module for selective cell lysis, separating means to eliminate centrifugation, high surface area pillar chip modules to purify DNA from a cell lysate, and microfluidic circuitry to integrate the steps in an automated platform.
1. A method of selectively lysing a specific type of cell, the method compnsing:
a. providing a sample to a self-contained, automated system, the sample includes at least two different cell types, wherein a first cell type is lysed at a first sonication energy;
b. automatically applying the first sonication energy to the sample by lysing means within the automated system thereby lysing the first cell type to form a first lysate and leaving intact one or more non-lysed cell types;
c. automatically separating the first lysate from the sample including the intact one or more non-lysed cell types by separating means within the automated system, thereby forming a separated sample including the intact one or more non-lysed cell types;
d. automatically moving the separated sample to the lysing means by a back-flow capability and microfluidic circuitry within the automated system, wherein the microfluidic circuitry includes the back-flow capability for moving the separated sample from the separating means to the lysing means; and
e. automatically lysing a second cell type from the separated sample by lysing means within the automated system thereby lysing the second cell type to form a second lysate.
2. The method of claim 1 further comprising automating a protein purification of the first lysate.
3. The method of claim 1 further comprising automating a protein purification of the second lysate.
4. The method of claim 1 wherein providing the sample is automated.
5. The method of claim 1 wherein the second cell type is lysed at a second sonication energy and automatically lysing the second cell type comprises automatically applying the second sonication energy to the sample.
6. The method of claim 5 wherein automatically lysing the second cell type further comprises automatically adding an additive to the sample prior to automatically applying the second sonication energy.
7. The method of claim 1 wherein automatically lysing the second cell type comprises automatically applying a chemical treatment to the second cell type.
8. The method of claim 1 further comprising automatically adding an additive to the sample prior to automatically applying the first sonication energy.
9. The method of claim 1 wherein the sample includes N additional different cell types, each additional cell type is automatically lysed at a different sonication energy, wherein N is an integer greater than or equal to one.
10. The method of claim 9 further comprising automatically applying a particular sonication energy to lyse a particular cell type and automatically separating the resulting lysate from the sample for each N additional different cell types, thereby forming N additional lysates.
11. The method of claim 9 wherein each successive application of sonication energy to the sample utilizes a sonication energy that is higher than the previous application.
12. The method of claim 9 further comprising automatically adding an additive to the sample prior to one or more applications of the sonication energy.
13. The method of claim 1 wherein automatically separating the first lysate from the sample comprises using a filter that passes the first lysate and blocks the second cell type.
14. The method of claim 1 wherein automatically separating the first lysate from the sample comprises automatically performing centrifugation on the sample to form a second cell type pellet and removing the first lysate.
15. The method of claim 1 wherein providing a sample comprises providing a sample matrix and applying the first sonication energy to the sample matrix to release the sample from the sample matrix.