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 particle and intracellular organelle alignment liquid (PIOAL) for use in a combined viscosity and geometric hydrofocusing analyzer, the PIOAL directing flow of a blood sample fluid of a given viscosity that is injected into a narrowing flowcell transition zone of the visual analyzer so as to produce a sample fluid stream enveloped by the PIOAL, the PIOAL comprising:
a fluid having a higher viscosity than the viscosity of the blood sample fluid,
a pH adjusting agent, and
sodium sulfate, sodium chloride, Procaine HCl, potassium phosphate monobasic, sodium phosphate dibasic, disodium EDTA, and deioinzed water,
wherein the pH adjusting agent is sodium hydroxide,
wherein a viscosity hydrofocusing effect induced by an interaction between the PIOAL fluid and the sample fluid associated with the viscosity difference, in combination with a geometric hydrofocusing effect induced by an interaction between the PIOAL fluid and the sample fluid associated with the narrowing flowcell transition zone, is effective to provide a target imaging state in at least some of the plurality of particles at an imaging site of the visual analyzer while a viscosity agent in the PIOAL 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
wherein the viscosity agent of the sheath fluid comprises glycerol at a concentration between about 1 to about 50% (v/v).
2. The PIOAL of claim 1 , wherein the viscosity agent of the sheath fluid comprises polyvinylpyrrolidone (PVP).
3. The PIOAL of claim 2 , wherein the polyvinylpyrrolidone (PVP) is at a concentration of 1% (w/v).
4. The PIOAL of claim 2 , wherein the viscosity agent of the sheath fluid further comprises glycerol.
5. The PIOAL 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% (w/v).
6. The PIOAL of claim 1 , wherein the PIOAL has a viscosity of between about 1-10 centipoise (cP).
7. The PIOAL of claim 1 , wherein the viscosity agent of the sheath fluid comprises glycerol at a concentration of about 30% (v/v).
8. The PIOAL of claim 1 , wherein the viscosity agent of the sheath fluid comprises glycerol at a concentration of about 6.5% (v/v).
9. The PIOAL of claim 1 , wherein the pH of the PIOAL is about 7.2.
10. The PIOAL of claim 1 , wherein the PIOAL is isotonic.
11. The PIOAL of claim 1 , further comprising a surfactant.
12. The PIOAL of claim 1 , further comprising an antimicrobial agent.
13. The PIOAL of claim 1 , wherein the PIOAL has a pH equal to the pH of the blood sample fluid.
14. A particle and intracellular organelle alignment liquid (PIOAL) for use in a combined viscosity and geometric hydrofocusing analyzer, the PIOAL directing flow of a blood sample fluid of a given viscosity that is injected into a narrowing flowcell transition zone of the visual analyzer so as to produce a sample fluid stream enveloped by the PIOAL, the PIOAL comprising:
a fluid having a higher viscosity than the viscosity of the blood sample fluid, and
a pH adjusting agent,
wherein the PIOAL consists of glycerol, sodium sulfate, sodium chloride, Procaine HCl, potassium phosphate monobasic, sodium phosphate dibasic, disodium EDTA, deionized water, and sodium hydroxide,
wherein a viscosity hydrofocusing effect induced by an interaction between the PIOAL fluid and the sample fluid associated with the viscosity difference, in combination with a geometric hydrofocusing effect induced by an interaction between the PIOAL fluid and the sample fluid associated with the narrowing flowcell transition zone, is effective to provide a target imaging state in at least some of the plurality of particles at an imaging site of the visual analyzer while a viscosity agent in the PIOAL 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
wherein the viscosity agent of the sheath fluid comprises glycerol at a concentration between about 1 to about 50% (v/v).