Method and apparatus for microfiltration to perform cell separation
A microfiltration apparatus and method for separating cells, such as circulating tumor cells, from a sample using a microfiltration device having a top porous membrane and a bottom porous membrane. The porous membranes are formed from parylene and assembled using microfabrication techniques. The porous membranes are arranged so that the pores in the top membrane are offset from the pores in the bottom membrane.
1. A method for capturing selected cells from a fluid sample comprising the cells, wherein the method comprises:
providing a microfiltration patch, wherein the microfiltration patch comprises:
a first membrane having several first membrane pores;
a second membrane having several second membrane pores, wherein the second membrane pores are distributed in the second membrane to be horizontally offset from the first membrane pores in the first membrane, and
a single gap defined by a planar space between a lower surface of the first membrane and an upper surface of the second membrane, wherein a distance between the lower surface of the first membrane and the upper surface of the second membrane is substantially even over the planar space, wherein the distance is smaller than diameters of the cells to be captured and wherein the single gap is further defined by a perimeter structure coupling the first membrane to the second membrane, and wherein no structures are present in the gap that obstruct the fluid sample flowing within the single gap from the first membrane pores to the second membrane pores, and the method further comprises:
applying the fluid sample to the first membrane of the microfiltration patch, and
flowing the fluid sample through the microfiltration patch to capture the selected cells.
2. The method according to claim 1 , wherein one or more of the first membrane pores have diameters smaller than diameters of the cells to be captured, whereby one or more of the cells to be captured become wedged in one or more of the several first membrane pores, and the locations of the first membrane pores and the second membrane pores and the distance between the lower surface of the first membrane and the upper surface of the second membrane are chosen such that a back pressure is applied to one or more of the wedged cells, wherein the back pressure is applied in a direction substantially opposite to a fluidic pressure applied to the one or more wedged cells.
3. The method according to claim 1 , wherein the first membrane pores have diameters larger than the cells to be captured.
4. The method according to claim 1 , wherein applying the fluid sample to the first membrane comprises applying the fluid sample at a pressure.
5. The method according to claim 1 , wherein the first membrane and/or the second membrane comprises parylene.
6. The method according to claim 1 , further comprising performing cell detection.
7. The method according to claim 6 , further comprising culturing captured cells for a chosen length of time.
8. The method according to claim 3 , further comprising: separating the first membrane from the second membrane; and performing cell detection.
9. The method according to claim 1 , wherein the first membrane, the second membrane or both the first and second membrane comprise a material selected from one of the following materials: polyimide; polysiloxane; polyester; polyacrylate; cellulose; and polycarbonate.
10. The method according to claim 1 , further comprising: decoupling the first membrane from the second membrane at the perimeter structure.
11. The method according to claim 1 , wherein the selected cells comprise circulating tumor cells and wherein the first membrane pores have diameters larger than diameters of the circulating tumor cells and the distance between the first membrane and the second membrane is smaller than diameters of the circulating tumor cells.
12. The method according to claim 1 , wherein the second membrane pores are arranged in hexagon shape around each first membrane pore.