Sheath fluid systems and methods for particle analysis in blood samples
Aspects and embodiments of the instant disclosure provide a particle and/or intracellular organelle alignment agent for a particle analyzer used to analyze particles contained in a sample. An exemplary particle and/or intracellular organelle alignment agent includes an aqueous solution, a viscosity modifier, and/or a buffer.
1. A method for imaging a plurality of particles using a particle analysis system configured for combined viscosity and geometric hydrofocusing, the particles included in a blood fluid sample having a sample fluid viscosity, the method comprising:
flowing a sheath fluid along a flowpath of a flowcell, the sheath fluid having a sheath fluid viscosity that differs from the sample fluid viscosity by a viscosity difference in a predetermined viscosity difference range;
injecting the blood fluid sample into the flowing sheath fluid within the flowcell so as to provide a sample fluid stream enveloped by the sheath fluid;
flowing the sample fluid stream and the sheath fluid through a reduction in flowpath size toward an imaging site, such that a viscosity hydrofocusing effect induced by an interaction between the sheath fluid and the sample fluid stream associated with the viscosity difference, in combination with a geometric hydrofocusing effect induced by an interaction between the sheath fluid and the sample fluid stream associated with the reduction in flowpath size, is effective to provide a target imaging state in at least some of the plurality of particles at the imaging site while a viscosity agent in the sheath fluid retains viability of cells in the sample fluid stream leaving structure and content of the cells intact when the cells extend from the sample fluid stream into the flowing sheath fluid; and
imaging the plurality of particles at the imaging site.
2. The method of claim 1 , wherein the target imaging state comprises a target orientation of one or more target particles in the flow relative to a focal plane of an imaging device used to acquire images at the imaging site.
3. The method of claim 1 , wherein the plurality of particles comprise a member selected from the group consisting of a red blood cell, a white blood cell, and a platelet.
4. The method of claim 1 , wherein the plurality of particles include a cell having an intraparticle structure comprising a member selected from the group consisting of an intracellular structure, an organelle, and a lobe.
5. The method of claim 1 , wherein the sheath fluid has a viscosity between 1-10 centipoise (cP).
6. The method of claim 1 , wherein the predetermined viscosity difference has an absolute value within a range from about 0.1 to about 10 centipoise (cP).
7. The method of claim 1 , wherein the viscosity agent of the sheath fluid comprises a member selected from the group consisting of glycerol, glycerol derivative, ethylene glycol, propylene glycol (dihydroxypropane), polyethylene glycol, polyvinylpyrrolidone (PVP), carboxymethylcellulose (CMC), water soluble polymer(s) and dextran.
8. The method of claim 1 , wherein the viscosity agent of the sheath fluid comprises glycerol at a concentration between about 1 to about 50% (v/v).
9. The method of claim 1 , wherein the viscosity agent of the sheath fluid comprises glycerol and polyvinylpyrrolidone (PVP).
10. The method of claim 1 , wherein the viscosity agent of the sheath fluid comprises glycerol at a concentration of 5% (v/v) and glycerol and polyvinylpyrrolidone (PVP) at a concentration of 1% (v/v).
11. The method of claim 1 , wherein the plurality of particles comprise a set of non-spherical particles, the blood fluid sample has a direction of flow at the imaging site, and at least 90% of the set of non-spherical particles are aligned within 20 degrees from a plane substantially parallel to the direction of flow.
12. A system for imaging a plurality of particles in a blood fluid sample having a sample fluid viscosity, the system for use with a sheath fluid having a sheath fluid viscosity that differs from the sample fluid viscosity by a viscosity difference in a predetermined viscosity difference range, the system comprising:
a flowcell having a flowpath and a sample fluid injection tube, the flowpath having a reduction in flowpath size;
a sheath fluid input in fluid communication with the flowpath of the flowcell so as to transmit a flow of the sheath fluid along the flowpath of the flowcell;
a blood fluid sample input in fluid communication with the injection tube of the flowcell so as to inject a flow of the blood fluid sample into the flowing sheath fluid within the flowcell, such that as the sheath fluid and the sample fluid flow through the reduction in flowpath size and toward an imaging site, a viscosity hydrofocusing effect induced by an interaction between the sheath fluid and the sample fluid associated with the viscosity difference, in combination with a geometric hydrofocusing effect induced by an interaction between the sheath fluid and the sample fluid associated with the reduction in flowpath size, provides a target imaging state in at least some of the plurality of particles at the imaging site while a viscosity agent in the sheath fluid retains viability of cells in the sample fluid stream leaving structure and content of the cells intact when the cells extend from the sample fluid stream into the flowing sheath fluid; and
an imaging device that images the plurality of particles at the imaging site.
13. The system of claim 12 , wherein the target imaging state comprises a target orientation of one or more target particles in the flow relative to a focal plane of an imaging device used to acquire images at the imaging site.
14. The system of claim 12 , wherein the plurality of particles comprise a member selected from the group consisting of a red blood cell, a white blood cell, and a platelet.
15. The system of claim 12 , wherein the plurality of particles include a cell having an intraparticle structure comprising a member selected from the group consisting of an intracellular structure, an organelle, and a lobe.
16. The system of claim 12 , wherein the predetermined viscosity difference has an absolute value within a range from about 0.1 to about 10 centipoise (cP).
17. The system of claim 12 , wherein the viscosity agent of the sheath fluid comprises a member selected from the group consisting of glycerol, glycerol derivative, ethylene glycol, propylene glycol (dihydroxypropane), polyethylene glycol, polyvinylpyrrolidone (PVP), carboxymethylcellulose (CMC), water soluble polymer(s) and dextran.
18. The system of claim 12 , wherein the viscosity agent of the sheath fluid comprises glycerol at a concentration between about 1 to about 50% (v/v).
19. The system of claim 12 , wherein the viscosity agent of the sheath fluid comprises glycerol and polyvinylpyrrolidone (PVP).
20. The system of claim 12 , wherein the viscosity agent of the sheath fluid comprises glycerol at a concentration of 5% (v/v) and glycerol and polyvinylpyrrolidone (PVP) at a concentration of 1% (w/v).
21. The system of claim 12 , wherein the plurality of particles comprise a set of non-spherical particles, the blood fluid sample has a direction of flow at the imaging site, and at least 90% of the set of non-spherical particles are aligned within 20 degrees from a plane substantially parallel to the direction of flow.