Hematology systems and methods
Inventors
Wanders, Bart J. • Farrell, Gregory A. • Adams, Thomas H. • Groner, Warren • Zhao, Xiaodong • Jordan, Brett • Cremins, Jack • Quon, Carol
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9702806-B2
Publication Date
2017-07-11
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. Embodiments also encompass systems, compositions, and methods for analyzing a sample containing particles. Particles such as blood cells can be categorized and counted by a digital image processor. A digital microscope camera can be directed, for example using certain focusing techniques, into a flowcell defining a symmetrically narrowing flowpath in which the sample stream flows in a ribbon flattened by flow and viscosity parameters between layers of sheath fluid. Blood cell images can be collected and analyzed using dynamic range extension processes and systems.
Core Innovation
The invention relates to hematology particle imaging systems and methods configured for combined viscosity hydrofocusing and geometric hydrofocusing. A sheath fluid is flowed along a flowpath of a flowcell and has a sheath fluid viscosity that differs from the sample fluid viscosity by a viscosity difference in a predetermined viscosity difference range. The blood fluid sample is injected into the flowing sheath fluid such that a sample fluid stream is enveloped by the sheath fluid.
The sheath fluid and sample fluid are flowed through a reduction in flowpath size toward an imaging site to induce a viscosity hydrofocusing effect and a geometric hydrofocusing effect. The combined focusing 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 and leaves structure and content intact as cells extend from the sample fluid stream into the flowing sheath fluid.
The invention further includes imaging the plurality of particles at the imaging site, including reorientation of cells so that a second subset is more numerous than a first subset based on major surfaces being oriented transverse to an orientation of an imaging path. The disclosed invention also describes flowing cells suspended in a viscosity agent within an alignment liquid so that non-spherical particles in a ribbon-shaped sample stream are aligned within a defined range relative to the direction of flow for improved imaging conditions for automated cytology based on image quality.
The document further describes particle types including RBCs, reticulocytes, nucleated RBCs, WBCs and WBC subtypes, and platelets. In the imaging workflow, improved alignment and focal placement support automated imaging-based analysis such as WBC differentials and morphological abnormality detection, with reduced manual review.
Claims Coverage
The independent claims cover five inventive features across method and system embodiments: combined viscosity and geometric hydrofocusing imaging with cell viability, cell subset reorientation, four-second-or-less delivery using processor-controlled injection, and imaging-based ratio calculation with a hematology cell counter.
Combined viscosity and geometric hydrofocusing for target imaging state with viable cells
Flowing a sheath fluid along a flowpath of a flowcell where sheath viscosity differs from sample viscosity by a viscosity difference in a predetermined viscosity difference range; injecting the blood fluid sample into the flowing sheath fluid to provide a sample fluid stream enveloped by the sheath fluid; flowing the sample fluid stream and sheath fluid through a reduction in flowpath size toward an imaging site so that viscosity hydrofocusing induced by viscosity difference, in combination with geometric hydrofocusing induced by reduction in flowpath size, provides a target imaging state while a viscosity agent retains viability of cells and leaves structure and content intact as cells extend into the flowing sheath fluid; imaging the plurality of particles at the imaging site.
Cell subset reorientation based on major surface orientation transverse to imaging path
The imaging step comprises imaging along an imaging path while the plurality of cells include a first subset with major surfaces oriented transverse to an orientation of the imaging path and a second subset with major surfaces oriented transverse to the imaging path, where the second subset is more numerous than the first subset; the interaction between the sheath fluid and the blood fluid sample associated with differing viscosities reorients at least some of the plurality of cells such that the second subset is more numerous than the first subset.
Delivering cells to imaging site in four seconds or less using combined focusing and sample flow rate
The viscosity hydrofocusing effect in combination with the geometric hydrofocusing effect and a flow rate of the blood fluid sample is effective to deliver cells in the blood fluid sample from a sample fluid injection tube to the imaging site in four seconds or less.
Hematology cell counter population and image-derived ratio for cell quantity measure
Acquiring images of a first number of a first cell type and a second number of a second cell type; determining a population of the second cell type in a second volume of the obtained sample by flowing the second volume through a hematology cell counter; determining a ratio of the first number to the second number using the acquired images; and calculating a cell quantity measure of the first cell type in the obtained sample using the ratio and the population of the second cell type.
Processor-controlled delivery to image capture site within four seconds or less
A processor coupled with the sample fluid injector system and the image capture device is configured to initiate injection at a sample flow rate so that the viscosity difference between sheath and blood fluid samples, in combination with the decrease in flowpath size and the flow rate of the sample, is effective to deliver cells from the sample fluid injection tube to the image capture site in four seconds or less, while a viscosity agent retains viability and leaves structure and content intact as cells travel to the image capture site.
Across the independent claims, the core claim coverage centers on combined viscosity hydrofocusing and geometric hydrofocusing in a flowcell using a sheath fluid viscosity difference to establish a target imaging state for particles or cells while a viscosity agent retains viability and maintains cell structure and content. Several claims further require reorientation of cells into a more numerous subset defined by major surfaces transverse to the imaging path, and some claims integrate imaging with hematology cell counter signals to calculate cell quantity measures or specify delivery of cells to an imaging or capture site within four seconds or less.
Stated Advantages
Provides a target imaging state for at least some of the plurality of particles at the imaging site.
Retains viability of cells in the sample fluid stream.
Leaves structure and content of cells intact when cells extend from the sample fluid stream into flowing sheath fluid.
Reorients cells such that a second subset becomes more numerous than a first subset.
Enables delivery of cells to the imaging site in four seconds or less.
Enables calculation of a cell quantity measure of a first cell type using a ratio from acquired images and a population from a hematology cell counter.
Improved image quality for high-resolution imaging by aligning non-spherical particles within specified alignment metrics.
Intracellular structures are positioned closer to the focal plane or within depth of field to support imaging.
Enables automated image-based WBC differentials.
Supports morphological abnormality detection.
Reduced manual review.
Documented Applications
Hematology particle imaging of a plurality of cells from a blood fluid sample using a particle analysis system configured for combined viscosity and geometric hydrofocusing.
Hematology-style cell quantity measurement by imaging cell types and using a hematology cell counter to determine population and calculate a cell quantity measure.
Automated image-based WBC differential analysis using improved aligned ribbon-shaped sample streams and focal placement of intracellular structures.
Detection of morphological abnormalities using the improved image quality from aligned cells.
Interested in licensing this patent?