Particle imaging systems and methods using acoustic radiation pressure

1. A method for imaging acoustically manipulated particles in an acoustic flow cytometer, comprising:

introducing a fluid containing a population of particles therein to a flow cell;

applying acoustic radiation pressure to the flow cell;

acoustically focusing the population of particles within the flow cell to concentrate the population of particles;

interrogating some of the concentrated population of particles with an interrogation source to obtain an optical signal from some of the population of particles to yield population statistical data; and

constructing a sampling matrix of particles from subpopulations of the concentrated population of particles to produce a high content image representative of the population of particles.

2. The method of claim 1 , further comprising correlating the population statistical data with the high content image to produce improved data content.

3. The method of claim 1 , further comprising slowing a transit time of some of the population of particles during the interrogating step.

4. The method of claim 1 , wherein at least a portion of the population of particles are labeled with one or more non-fluorescent absorptive dyes.

5. The method of claim 4 , wherein the one or more non-fluorescent absorptive dyes comprise different colors and concentrations.

6. The method of claim 1 , wherein the interrogation source is a light source, and wherein the subpopulations of the concentrated population of particles are gated based on one or more of a scatter signature and a fluorescence signature.

7. The method of claim 6 , further comprising creating light scatter along with one or more wavelengths for analyzing the particles.

8. The method of claim 1 , wherein at least a portion of the population of particles comprises white blood cells.

9. A method of acoustically focusing particles in a plane, comprising:

transiting a fluid containing particles therein through a channel at a flow rate;

adjusting the flow rate for a desired transit time through an interrogation zone through which a light from an excitation source passes;

optically exciting the particles with the excitation source;

detecting an optical signal from the particles; and

analyzing the optical signal.

10. The method of claim 9 , further comprising disposing a substantially acoustically transparent gas-contacting membrane at a top surface of the channel wherein a pressure node is located outside of the channel at a gas interface.

11. The method of claim 9 , further comprising imaging the particles.

12. The method of claim 9 , further comprising creating light scatter along with one or more wavelengths for analyzing the particles.

13. The method of claim 9 , wherein the particles are microsphere beads.

14. The method of claim 9 , wherein the particles are cells, and further comprising imaging the cells, wherein imaging the cells comprises capturing cellular morphology, orientation, and internal structure.

15. The method of claim 9 , wherein the fluid is a reagent.

16. An acoustic flow cytometer capable of measuring axial light loss, comprising:

a channel having an inlet for accepting a fluid sample stream of analytes;

an acoustic signal producing transducer coupled to the channel wherein the acoustic signal producing transducer produces an acoustic signal to the channel to induce within the channel acoustic radiation pressure to concentrate the analytes within the fluid sample stream;

optical equipment for analyzing the analytes wherein the optical equipment comprises a light source or sources with one or more wavelengths for analyzing the analytes when the analytes pass through the one or more wavelengths; and

at least one linear array detector configured to increase signal by spatial isolation of axial light loss to detect absorbance from the analytes to determine characteristics of the analytes.

17. The acoustic flow cytometer of claim 16 , wherein the at least one linear array detector is configured to provide information regarding both axial light loss and a plurality of angles of light scatter.

18. The acoustic flow cytometer of claim 16 , wherein the acoustic flow cytometer is configured to slow a transit time of the analytes while the analytes pass through the one or more wavelengths, and wherein at least a portion of the analytes are labeled with one or more non-fluorescent absorptive dyes having different colors and concentrations.

19. The method of claim 1 , further comprising disposing a substantially acoustically transparent gas-contacting membrane at a top surface of the flow cell wherein a pressure node is located outside of the flow cell at a gas interface.

20. The method of claim 8 , wherein the portion of the population of particles comprising white blood cells further comprises red blood cells, and wherein the method further comprises immunophenotyping at least some of the white blood cells without performing red blood cell lysis.