Compositions and methods for disease diagnosis using single cell analysis
Certain embodiments of the invention are directed to evaluating and identifying cells by recording and interpreting a time-dependent signal produced by unique cell respiration and permeability attributes of isolated viable cells.
1. A method for identifying a cell comprising:
(a) contacting a sample containing cells with at least one reporter forming a sample mixture;
(b) partitioning the sample mixture into a plurality of small volume partitions, wherein some small volume partitions contain one cell or one cellular aggregate;
(c) incubating the small volume partitions for a period of time at a specified temperature or series of temperatures;
(d) monitoring optical characteristics of the small volume partitions during the incubation time;
(e) constructing a waveform for each small volume partition based on the optical characteristics over time; and
(f) identifying the cell in the small volume partition using information provided by analysis of the waveform for the particular small volume partition.
2. The method of claim 1 , wherein the small volume partitions are droplets in an immiscible fluid.
3. The method of claim 1 , further comprising arranging the small volume partitions in a two-dimensional array.
4. The method of claim 3 , wherein the two-dimensional array is a static two-dimensional array.
5. The method of claim 2 , wherein the immiscible fluid is a fluorocarbon comprising a fluorosurfactant.
6. The method of claim 2 , wherein partitioning of the sample is done by Laplace pressure gradients.
7. The method of claim 2 , wherein partitioning of the sample is done by using shear stress.
8. The method of claim 1 , wherein monitoring the optical characteristics of the small volume partitions is performed using a camera.
9. The method of claim 1 , wherein the at least one reporter is a fluorescent dye.
10. The method of claim 9 , where the reporter is a resazurin dye.
11. The method of claim 9 , where the at least one reporter is tetrazolium dye.
12. The method of claim 9 , wherein the monitoring optical characteristics of the small volume partitions further comprises exciting the reporter using a light emitting diode (LED).
13. The method of claim 1 , wherein the incubating of the small volume partitions is at a constant temperature.
14. The method of claim 13 , wherein the temperature is 37° C.
15. The method of claim 1 , wherein the cell is a microbe.
16. The method of claim 15 , further comprising identifying the microbe by species, genus, family, order, class, phylum, kingdom, or a combination thereof.
17. The method of claim 15 , wherein the micorobe is a bacterium.
18. The method of claim 17 , wherein classifying the bacterium is by gram-stain group.
19. The method of claim 1 , wherein the cell is a eukaryotic cell.
20. The method of claim 1 wherein the identification of the cell using the waveform includes comparing the small volume partition waveform to a library of stored waveforms.