Cytocolor, Inc.

Other

International Remote Imaging Systems, Inc.

Lawrence Kass

Leonard Spolter

A composition and method for separating red blood cells from whole blood comprising a rouleaux-forming aggregator and an enhancer for enhancing the settling rate. The enhancer is a material which alters directly or indirectly the properties of the red blood cell and may alter the structure and/or reactivity of the aggregator, without adversely affecting the morphology and function of white blood cells. In most instances, the red blood cell enhancers of the invention are osmotic agents. Such agents create a hypertonic solution while not entering the cells themselves. Preferably the enhancer is a salt of oxalic acid, a salt of malonic acid, mannitol or sucrose. Potassium oxalate is most preferred. High molecular weight substances which are large enough to form molecular bridges between red blood cells form the aggregators used in the invention. Examples of useful aggregators include dextran, hespan, pentaspan, ficoll, gum arabic, polyvinylpyrrolidone, nucleic acids, and proteins such as fibrinogen and gamma globulins.

Composition and method for enrichment of white blood cells from whole

Peripheral blood lymphocytes displayed a plurality of sizes and colors when exposed first to a fixative solution of C.I. basic blue 141, then to an aqueous alkaline solution of the same dye and subsequently treated with a neutral HEPES buffer solution containing small but effective amounts of various metal compounds. As confirmed with purified lymphocyte subpopulations obtained with a cell sorter, T helper cells (CD4) were small and their nuclei and cytoplasm stained deep blue. T cytotoxic/suppressor cells (CDS) were larger than T helper cells, their nuclei stained pale green or blue green and their cytoplasm contained a cluster of magenta colored granules. Used in the process described, basic blue 141 provides means of identifying and differentiating CD4 and CD8 cells under the ordinary light microscope without using monoclonal antibodies or fluorescence.

Oxazine stained lymphocytes and method

A composition and method for separating red blood cells from whole blood including a rouleaux-forming aggregator and an enhancer for enhancing the settling rate. The enhancer is a material which alters directly or indirectly the properties of the red blood cell and may alter the structure and/or reactivity of the aggregator, without adversely affecting the morphology and function of white blood cells. In most instances, the red blood cell enhancers of the invention are osmotic agents. Such agents create a hypertonic solution while not entering the cells themselves. Preferably the enhancer is a salt of oxalic acid, a salt of malonic acid, mannitol or sucrose. Potassium oxalate is most preferred. High molecular weight substances which are large enough to form molecular bridges between red blood cells form the aggregators used in the invention. Examples of useful aggregators include dextran, hespan, pentaspan, ficoll, gum arabic, polyvinylpyrrolidone, nucleic acids, and proteins such as fibrinogen and gamma globulins.

This invention is directed to the cytology of blood, bone marrow and lymph node cells and to the method of differentiating, identifying and enumerating monocytes among a plurality of cells of hematopoietic origin. More specifically, the invention is directed to stained cells and to the use of a cationic sulfur-containing azo dye capable of staining monocytes a bright red-violet color. The dye is a water soluble quaternary azo dye identified in the Colour Index as Basic Blue 54 (C.I. 11052). The stained monocytes obtained by the process of this invention have color characteristics which permit the identification and differentiation of said cells by use of various instruments including, for example, an image analyzer, microscope, photo microscope, and the like.

Method of staining monocytes and compositions thereof

This invention relates to the cytology of blood, bone marrow, and lymph node cells, and to the composition and method of differentiating, identifying, and enumerating said cells among a plurality of cells of hematopoietic origin. More specifically, the invention is directed to the use of basis cationic diazo dyes capable of staining a plurality of cells of hematopoietic origin to differentiate and enumerate the abnormal from the normal cells. The stained cells obtained by this invention have individual color characteristics which permit the identification and differentiation by use of various optical instruments, including, for example, an image analyzer, microscope, photomicroscope and the like.

Cells stained with a diazo dye

This invention relates to the cytology of blood, bone marrow, and lumpth node cells, and a method of differentiating, identifying, and enumerating said cells among a plurality of cells of hematopoietic origin. More specifically, the invention is directed to the use of basic cationic diazo dyes capable of staining a plurality of cells of hematopoietic origin to differentiate and enumerate the abnormal from the normal cells. The stained cells obtained by this invention have individual color characteristics which permit the identification and differentiation by use of various optical instruments, including, for example, an image analyzer, microscope, photomicroscope and the like.

Method of staining cells with a diazo dye and compositions thereof

This invention is directed to the use of specific polymethine dyes for staining fixed biopsy specimens of hematopoietic origin consisting of blood, bone marrow and lymph node cells for purposes of identifying and enumerating the stained cells when viewed under dark field illumination. The polymethine dyes, e.g. Astrazon Red Violet FRR, stain the lymphocyte subpopulations distinct colors capable of being viewed under dark field illumination for purposes of distinguishing one stained cell from another. This method of staining and the compositions thereof represents an advance in the art of lymphocyte subpopulation identification which was achieved heretofore primarily by using monoclonal antibody techniques.

Method of identifying polymethine stained lymphocyte subpopulations and

This invention is directed to the cytology of blood, bone marrow and lymph node cells and the method of differentiating, identifying and enumerating said cells among a plurality of cells of hematopoietic origin. More specifically, the invention is directed to the use of a single cationic sulfur-containing azo dye in solution capable of staining said cells in a fixative. The dye is a water soluble quaternary azo dye identified in the Color Index as Basic Blue 41 capable of staining a plurality of cells including malignant cells of hematopoietic origin wherein the stained cells acquire individual color characteristics permitting the differentiation, identification and enumeration of each stained cell. The stained cells obtained by the process of this invention have individual color characteristics which permit the identification and differentiation by use of various instruments including, for example, an image analyzer, microscope, photomicroscope, and the like.

Method of staining and identifying cells and compositions thereof

Differentiation, identification and enumeration of sub-populations of lymphocytes including B-cells and T-cells can now be further discriminated into their sub-populations by means of basic orange 21 as a supravital stain to provide distinctive morphologic differences between B-cells, T-suppressor cells, T-helper cells and natural killer (NK) cells without resorting to the use of individual monoclonal antibodies for differentiation of each of the above. The differentiation, identification and enumeration of the foregoing cells can be accomplished by means of both white light absorbance and fluorescence measurements, singly or in sequential combinations.

Metachromatic dye sorption means for differential determination of

Lengthy cytochemical procedures for differentiation, identification, enumeration and diagnostic study of leukocytes in human blood have been advanced and shortened by a rapid method of optical differentiation of the five individual white blood cell species by selective use of basic quaternary metachromatic dye staining of supravital blood at controlled temperature. Presently a manual method, the improvement provides essential groundwork for improved accuracy in automated differential leukocyte counting.

Individual leukocyte determination by means of differential

Differentiation, identification and enumeration of dye responsive human blood cells essential to diagnosis and prognosis of certain diseases is established by preparation of the reaction product of a human blood specimen of the patient in an aqueous fixative-free environment at blood temperatures with selected basic, cationic quaternary dyes, particularly basic orange #21. The so-prepared specimen becomes instantly responsive to emissive wave energy source exposure to stimulate reproducible fluorescent light emission from all of said dye responsive cells. The quantum of said fluorescent light energy emitted provides individual sets of data response relative to hue, value, chroma and intensity to permit differentiation, identification and enumeration of all of said dye responsive components in said blood specimen. By use of a bi-modal wave energy system involving additional use of white light wave energy absorbance, independent sets of data can provide both confirmatory and ancillary information.

Metachromatic dye sorption and fluorescent light emmisive means for

In the diagnosis and prognosis of various diseases as well as medical researches in relation thereto studies leading to differentiation, identification, enumeration and following the variations in human blood, leukocytes play a vital role. Heretofore, accurate detailed lengthy and arduous procedures were essential to such ends, if available. In a parent case, of which this is a continuation-in-part, it was found selective use of certain basic quaternary, metachromatic dye staining by supravital techniques made possible instantaneous optical differentiation of the five major individual leukocyte species. Here further research has revealed that with but one of the species of the dye class above one can by the same techniques make the foregoing determinations with regard to specific leukocytes including promyelocytes, myelocytes, metamyelocytes, band cells, T-cells and B-cells as well as the neutrophils, eosinophils, basophils and monocytes of the parent disclosure.

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Hinckley, OH, United States of America

Lawrence Kass