Apparatus and methods for conducting assays and high throughput screening
The present invention provides microfluidic devices and methods for using the same. In particular, microfluidic devices of the present invention are useful in conducting a variety of assays and high throughput screening. Microfluidic devices of the present invention include elastomeric components and comprise a main flow channel; a plurality of branch flow channels; a plurality of control channels; and a plurality of valves. Preferably, each of the valves comprises one of the control channels and an elastomeric segment that is deflectable into or retractable from the main or branch flow channel upon which the valve operates in response to an actuation force applied to the control channel.
1. A method of cell analysis comprising
(a) introducing a solution comprising eukaryotic cells into a microfluidic device that comprises holding spaces in which cells may be retained, so that in a plurality of said holding spaces a single cell is retained;
(b) within each said holding space, contacting the cell with an agent that causes the cell to lyse thereby producing a cell lysate; and
(c) carrying out an assay with a component of the cell lysate, wherein the assay is carried out in the microfluidic device.
2. The method of claim 1 wherein the assay is a binding assay in which the antiligand is an antibody and the ligand is an antigen recognized by the antibody.
3. The method of claim 1 wherein the assay comprises a primer extension reaction.
4. The method of claim 3 wherein the primer extension reaction is template-directed DNA synthesis.
5. The method of claim 3 wherein the primer extension reaction comprises extension of a primer by the action of reverse transcriptase.
6. The method of claim 3 wherein primer extension reaction is step in a sequencing reaction.
7. The method of claim 3 wherein the primer extension reaction is a step in a polymerase chain reaction.