Microfluidic chamber assembly for mastitis assay
Inventors
Rodriguez, Rodolfo R. • Galanaugh, Charles F.
Assignees
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Publication Number
US-10983104-B2
Publication Date
2021-04-20
Expiration Date
Abstract
The present invention relates to a device and method for the detection of mastitis or other disease from a body fluid of a mammal for example from cow's milk. The device and method relates to a wedge microfluidic chamber for using a minimal amount of fluid and being able to use the device to observe leukocytes in a mono-layer for the purpose of disease detection, cell counts or the like.
Core Innovation
The invention provides a microfluidic wedge chamber assembly and associated methods for performing a leukocyte differential assay on a milk sample comprising live cells. The assembly includes a wedge base and a wedge top positioned so that the wedge top has a first top edge and an opposite second edge in an elevated configuration relative to the base. A cut out well is positioned to add the milk sample to the wedge, at least one of the top or base is optically transparent, and a leukocyte observation colorant is positioned within the chamber.
When the sample is received in the well, the milk sample flows through a void between the top and the base into the chamber via capillary action and forms a mono-layer of cells in the chamber. An automatic reader observes an image morphology of live cell leukocytes in the chamber, enumerates and calculates leukocytes in the milk-containing chamber into sub-populations, and derives a differential count of the leukocyte sub-populations.
The same microfluidic wedge chamber assembly architecture is also used for automated imaging of live cell leukocyte morphology in a milk sample, identification of leukocyte subpopulations based on image morphology, and derivation of a leukocyte-derived differential. The leukocyte-derived differential is used to determine a health status including a stage of disease comprising a stage of mastitis, including mastitis during the peripartum period and/or early lactation.
Claims Coverage
The document includes two independent claims. The claims center on a microfluidic wedge chamber assembly with an optically transparent wedge and leukocyte observation colorant that forms a monolayer and supports automatic reader imaging, enumeration, calculation, and identification of live-cell leukocyte sub-populations.
Microfluidic wedge chamber with optically transparent wedge and leukocyte observation colorant
A milk sample comprising live cells is added to a microfluidic chamber assembly comprising a wedge base and a wedge top positioned in an elevated configuration, with a cut out well for adding the sample, at least one of the top or base being optically transparent, and a leukocyte observation colorant positioned within the chamber. The sample flows through a void between the top and the base via capillary action and forms a mono-layer of cells in the chamber.
Automatic reader imaging, enumeration, sub-population calculation and differential count
An automatic reader observes an image morphology of live cell leukocytes in the chamber, enumerates and calculates leukocytes into sub-populations based on the observation, and derives a differential count of the leukocyte sub-populations.
Automated imaging and identification of leukocyte subpopulations
An automatic reader observes an image morphology of live cell leukocytes in the chamber and identifies leukocyte subpopulations based on the image morphology of the live cell leukocytes.
The inventive features combine a microfluidic wedge chamber that forms a monolayer of live cells via capillary flow with automatic reader-based imaging, enumeration, calculation, and identification of leukocyte sub-populations to derive a differential count and related disease-stage determination.
Stated Advantages
Enables an automated leukocyte differential assay performed in the automatic reader.
Provides differential counts of leukocyte sub-populations based on image morphology of live cell leukocytes.
Enables automated imaging of live cell leukocyte morphology and identification of leukocyte subpopulations from image morphology.
Supports determining a health status as a stage of disease comprising a stage of mastitis, including mastitis during the peripartum period and/or early lactation.
Documented Applications
Determining health status/stage of disease comprising a stage of mastitis, including mastitis during the peripartum period and/or early lactation.
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