Method and composition for staining and processing a urine sample
Inventors
Assignees
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Publication Number
US-9939356-B2
Publication Date
2018-04-10
Expiration Date
Abstract
The present disclosure relates to a staining methodology employing a particle contrast agent composition capable of rapidly staining cells in a single step. The particle contrast agent composition can be comprised of a combination of one or more particle contrast agents and one or more permeabilizing agents, optionally including one or more fixing agents and other components. The particle contrast agent composition can include Crystal Violet, 5PD-Lytic, and Proclin 300.
Core Innovation
The invention provides a particle contrast agent composition for staining particles of a urological fluid sample for imaging in an automated particle analysis system. The composition includes a particle contrast agent selected from Crystal Violet, New Methylene Blue, Safranin O, Eosin Y, and Methyl Green, together with a permeabilizing agent including a 5-part differential (5 PD) lytic reagent including saponin and an antimicrobial agent.
The described compositions and examples combine the selected particle dyes with the 5 PD lytic reagent containing saponin, with optional inclusion of a fixing agent and other solution constituents. Crystal Violet and Safranin O are used in particular embodiments, and phosphate buffered saline components and optionally sodium chloride are included in certain formulations, with fixing agents optionally selected from glutaraldehyde, formaldehyde, and diazolidinyl urea.
The invention also provides a one-step method of treating particles of a urological fluid sample for imaging in an automated particle analysis system. A particle contrast agent composition and the urological fluid sample are combined to form a sample mixture with a final concentration of the particle contrast agent composition by weight of the sample mixture between about 1% and about 20%, and the sample mixture is incubated at a temperature above 20°C for fewer than 90 seconds.
Claims Coverage
The patent includes two independent claims: a composition claim and a method claim. Both are centered on a defined particle contrast agent component, a 5 PD lytic reagent including saponin at specified staining-condition concentrations, and an antimicrobial agent; the method claim additionally specifies mixture concentration and incubation conditions.
Particle contrast agent composition for automated particle analysis imaging
A particle contrast agent composition for staining particles of a urological fluid sample for imaging in an automated particle analysis system comprising a particle contrast agent selected from Crystal Violet, New Methylene Blue, Safranin O, Eosin Y, and Methyl Green; a permeabilizing agent including a 5-part differential (5 PD) lytic reagent including saponin present in concentrations of about 0.01 mg/L to about 100 mg/L under staining conditions; and an antimicrobial agent.
Rapid treating method using concentration-limited particle contrast agent and short incubation
A method of treating particles of a urological fluid sample for imaging in an automated particle analysis system comprising combining a particle contrast agent composition and the urological fluid sample into a sample mixture resulting in a final concentration of the particle contrast agent composition by weight of the sample mixture between about 1% and about 20%; and incubating the sample mixture at a temperature above 20° Celsius for fewer than 90 seconds, wherein the particle contrast agent composition comprises a particle contrast agent selected from Crystal Violet, New Methylene Blue, Safranin O, Eosin Y, and Methyl Green; a permeabilizing agent including a 5 PD lytic reagent including saponin present in amounts sufficient to result in concentrations of about 0.01 mg/L to about 100 mg/L under staining conditions; and an antimicrobial agent.
Across the independent claims, the core inventive concept is a particle staining composition that pairs a selected particle contrast dye set with a 5 PD lytic reagent containing saponin at specified staining-condition concentrations and an antimicrobial agent, and a corresponding method that combines the composition with a urological fluid to achieve a defined final concentration and incubates the mixture above 20°C for fewer than 90 seconds.
Stated Advantages
Rapid, single-step particle staining/processing for automated urine particle imaging in an automated particle analysis system.
Facilitates visual distinction of particles, including RBC, WBC, epithelial cells, bacteria, yeast, casts, crystals, and trichomonas, for automated particle analysis/particle appearance outcomes.
Documented Applications
Staining particles of a urological fluid sample for imaging in an automated particle analysis system, including automated urine particle imaging using automated flow cytometry/flowcell imaging.
Treating particles of a urological fluid sample for imaging in an automated particle analysis system using a sample mixture incubated for fewer than 90 seconds.
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