Sheath fluid systems and methods for particle analysis in blood samples
Inventors
Farrell, Gregory A. • Wanders, Bart J. • Adams, Thomas H. • Groner, Warren • Zhao, Xiaodong
Assignees
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Publication Number
US-9316635-B2
Publication Date
2016-04-19
Expiration Date
Abstract
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.
Core Innovation
A particle and intracellular organelle alignment liquid (PIOAL) is provided for use in a combined viscosity and geometric hydrofocusing analyzer. The PIOAL directs flow of a blood sample fluid into a narrowing flowcell transition zone of a visual analyzer, producing a sample fluid stream enveloped by the PIOAL.
The PIOAL fluid includes a fluid having a higher viscosity than the viscosity of the blood sample fluid and a pH adjusting agent, together with specified constituents including sodium sulfate, sodium chloride, Procaine HCl, potassium phosphate monobasic, sodium phosphate dibasic, disodium EDTA, deionized water, and sodium hydroxide. The PIOAL 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.
The target imaging state results from a viscosity hydrofocusing effect induced by interaction between the PIOAL fluid and the sample fluid associated with the viscosity difference, combined with a geometric hydrofocusing effect induced by interaction between the PIOAL fluid and the sample fluid associated with the narrowing flowcell transition zone. 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.
Claims Coverage
The document includes two independent claims directed to a particle and intracellular organelle alignment liquid (PIOAL) for a combined viscosity and geometric hydrofocusing analyzer. Across the independent claims, the coverage centers on a higher-viscosity PIOAL at a narrowing flowcell transition zone, a combined viscosity hydrofocusing effect and geometric hydrofocusing effect to reach a target imaging state, and retention of cell viability with intact cell structure and content during extension into a flowing sheath fluid, with glycerol-based sheath-fluid viscosity specified.
Combined viscosity and geometric hydrofocusing PIOAL enveloping a blood sample stream
A PIOAL for use in a combined viscosity and geometric hydrofocusing analyzer that directs flow of a blood sample fluid into a narrowing flowcell transition zone of a visual analyzer so as to produce a sample fluid stream enveloped by the PIOAL, where the PIOAL comprises a fluid having a higher viscosity than the viscosity of the blood sample fluid and a pH adjusting agent.
Specific PIOAL formulation with sodium hydroxide pH adjusting agent
The PIOAL comprises a pH adjusting agent that is sodium hydroxide and includes sodium sulfate, sodium chloride, Procaine HCl, potassium phosphate monobasic, sodium phosphate dibasic, disodium EDTA, and deionized water.
Target imaging state via viscosity and geometric hydrofocusing effects while retaining viability
A viscosity hydrofocusing effect induced by interaction between the PIOAL fluid and the sample fluid associated with the viscosity difference, in combination with a geometric hydrofocusing effect induced by 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 and leaves structure and content of the cells intact when the cells extend into the flowing sheath fluid.
Glycerol-containing sheath-fluid viscosity agent concentration range
The viscosity agent of the sheath fluid comprises glycerol at a concentration between about 1 to about 50% (v/v).
PIOAL limited component set including glycerol and named salts
The PIOAL consists of glycerol, sodium sulfate, sodium chloride, Procaine HCl, potassium phosphate monobasic, sodium phosphate dibasic, disodium EDTA, deionized water, and sodium hydroxide.
Across both independent claims, the inventive coverage is directed to a higher-viscosity, pH-adjusted PIOAL that envelops a blood sample stream at a narrowing flowcell transition zone to generate a combined viscosity hydrofocusing effect and geometric hydrofocusing effect, thereby providing a target imaging state while a viscosity agent retains cell viability and preserves cell structure and content during extension into the flowing sheath fluid. The sheath-fluid viscosity agent is specified to include glycerol at about 1% to about 50% (v/v), and one independent claim further limits the PIOAL to a specific named component set.
Stated Advantages
Provides a target imaging state in at least some of the plurality of particles at an imaging site of the visual analyzer.
Retains viability of cells in the sample fluid stream.
Leaves structure and content of the cells intact when the cells extend into the flowing sheath fluid.
Documented Applications
Imaging of a plurality of particles at an imaging site of a visual analyzer using a combined viscosity and geometric hydrofocusing analyzer.
Cell/particle analysis use case for hematology particle analysis, including white blood cell (WBC) differentials.
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