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AssigneeRobert A. Levine
InventorsDavid L. Rimm, Robert A. Levine, Stephen C. Wardlaw, Paul Fiedler
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A method for analyzing blood enables one to isolate, detect, enumerate and confirm under magnification the presence or absence of target cancer cells and/or hematologic progenitor cells which are known to circulate in blood. The analysis is performed in a sample of centrifuged anticoagulated whole blood. The analysis involves both morphometric and epitopic examination of the blood sample while the blood sample is disposed in a centrifuged blood sampling tube. The epitopic analysis of the presence or absence of cancer cells relies on the detection of epitopes which are known to present only on cancer cells; and the epitopic analysis of the presence or absence of hematologic progenitor cells relies on the detection of epitopes which are known to present only on hematologic progenitor cells. The targeted epitopes on the target cell types are epitopes which are also known to be absent on normal circulating blood cells; and the target cancer cell epitopes are epitopes which are known to be absent on target hematologic progenitor cells. Fluorophors with distinct emissions are coupled with antibodies which are directed against the targeted epitopes. The morphometric analysis is performed by staining the cells in the blood sample with an intracellular stain such as acridine orange which highlights the intracellular cell structure. Both the morphometric and epitopic analyses are preferably performed at or near the platelet layer of the expanded buffy coat in the centrifuged blood sample. The morphometric analysis and/or the epitopic analysis may be performed under magnification both visually and/or photometrically.
A method for detecting the presence or absence of individual circulating epithelial cancer cells in an anticoagulated whole blood sample, said method comprising the steps of:
A method for detecting the presence or absence of cancerous cell morphology in individual circulating epithelial cells in a centrifuged sample of anticoagulated whole blood contained in a container which container also contains an insert which forms a free volume in the container, said blood sample having been combined with one or more labeling agents that are specific to one or more epitopes on the epithelial cells, and said blood sample having also been combined with a colorant which is operable to clarify epithelial cell morphology in individual epithelial cells in the blood sample, said method comprising the steps of identifying a percentage of all individual labeled epithelial cells which are disposed in said free volume in situ in the container, and examining the cell morphology of any such individual identified epithelial cells in situ in the container so as to determine whether any such individual identified epithelial cells display cancerous cell morphology.
A method for identifying individual circulating cancerous epithelial cells in a centrifuged sample of anticoagulated whole blood which sample is contained in a transparent container, which container also contains an insert, and which blood sample has been combined with at least one labeling agent that is specific to at least one epithelial cell epitope, and which blood sample has also been combined with a colorant that clarifies nucleated cell morphology, said method comprising the steps of examining a free volume located between the insert and the container wall wherein buffy coat constituents in the blood sample gravitate during centrifugation and enumerating any individual labeled epithelial cells having cancerous morphology in situ in the container, which individual labeled epithelial cells have localized during centrifugation in said free volume in the container.
A method for differentiating individual cancer cells from individual hematologic progenitor cells and from other nucleated cells in a sample of anticoagulated whole blood, said method comprising the steps of:
A method for detecting individual cancer cells and/or individual hematologic progenitor cells in a sample of anticoagulated whole blood, said method comprising the steps of:
A method for analyzing a sample of anticoagulated whole blood in order to determine the presence or absence of individual cancer cells in the sample, said method comprising the steps of:
A method of identifying individual circulating epithelial cancer cells in a centrifuged sample of anticoagulated whole blood which sample is contained in a transparent container, which container also contains an insert that forms a well defined free volume in the container, and which blood sample has been combined with at least one labeling agent that is specific to at least one epithelial cancer cell epitope, said method comprising the steps of examining said well-defined free volume in the container wherein white cells and platelets in the blood sample have gravitated during centrifugation;
A method for detecting the presence or absence of individual circulating nucleated epithelial cells in an anticoagulated whole blood sample, said method comprising the steps of:
A method for detecting the presence or absence of individual circulating hematologic progenitor nucleated cells in an anticoagulated whole blood sample, said method comprising the steps of:
A method for detecting the presence or absence of individual circulating cancer cells in a centrifuged sample of anticoagulated whole blood contained in a container which container also contains an insert that forms a well-defined free volume in the container, said blood sample having been combined with one or more epitope-specific labeling agents that are operative to produce a characteristic signal result on individual cancer cells, and which result is defined by the presence of one or more epitopes on the individual cancer cells, and said blood sample having also been combined with a colorant which is operable to clarify cell morphology in all nucleated cells in the blood sample, said method comprising the steps of identifying by cell morphology all nucleated cells disposed in said well-defined annular free volume, and further characterizing all identified individual nucleated cells as cancer cells or non-cancer cells epitopically, said identifying and characterizing steps being performed in situ in the container.
Dependencies are inferred from the claim text.