Sysmex Corporation

Toa Medical Electronics Co., Ltd.

Tao Medical Electronics Co., Ltd.

Yukio Hamaguchi

Berend Houwen

Takashi Sakata

Takashi Morikawa

Yukio Tsujino

Yoshiro Ikeuchi

Fu-sheng Wang

Tomohiro Tsuji

Kinya Uchihashi

Hiroyuki Fujimoto

Yusuke Mori

Atsushi Shirakami

Rolf Mast

Teruya Matsumoto

Kenji Ito

Kazuhiro Moriyama

Ikuya Takenaka

Hitomi Ohmi

Hiroshi Okada

Kojiro Hirai

Syouzou Toda

A method for classifying and counting nucleated bone marrow cells comprises the steps of: (1) mixing a sample of bone marrow fluid with an erythrocyte lysing agent to lyse erythrocytes in the sample and render leukocytic cells and erythroid cells in the sample suitable for staining, and staining the sample with a fluorescent dye for producing a difference in intensity of fluorescence between the leukocytic cells and the erythroid cells; (2) introducing the resulting sample to a flow cytometer to detect at least one kind of scattered light and at least one kind of fluorescence; (3) classifying and counting nucleated bone marrow cells, the leukocytic cells and the erythroid cells with use of a difference in the intensity of the fluorescence and the scattered light; (4) calculating the ratio of the nucleated bone marrow cells to the erythroid cells or leukocytic cells from the obtained erythroid cell count or leukocytic cell count and the obtained nucleated bone marrow cell count; and (5) calculating the ratio of the leukocytic cells to the erythroid cells from the erythroid cell count and the leukocytic cell count.

Method of classifying and counting nucleated bone marrow cells

Improved method for detecting and/or counting the appearance of hematopoietic progenitor cells which comprises the steps of mixing a blood sample with a reagent capable of detecting immature cells without employing any immunological techniques, obtaining cell information about the thus treated sample using a particle analyzer and constructing a cell distribution profile, delineating a portion of the profile as a zone in which at least one subclass of hematopoietic progenitor cells appear, and counting the cells within the zone. The method is capable of detecting the appearance of hematopoietic progenitor cells in a simple and positive manner.

Method of detecting hematopoietic progenitor cells

A method for discriminating and counting erythroblasts comprises the steps of: (i) staining leukocytes in a hematologic sample by adding a fluorescent labeled antibody capable of binding specifically with leukocytes to the hematologic sample; (ii) raising the permeability only of cell membranes of erythroblasts in the hematologic sample to a nucleotide fluorescent dye which does not permeate a cell membrane usually, the nucleotide fluorescent dye having a fluorescent spectrum capable of being distinguished from that of a fluorescent labeling compound of the fluorescent labeled antibody in step (i); (iii) staining nuclei of the erythroblasts in the hematologic sample with the nucleotide fluorescent dye; (iv) subjecting the hematologic sample to flowcytometry to detect at least two fluorescent signals from each cell; and (v) discriminating and counting the erythroblasts from difference in intensity between the at least two fluorescent signals.

Process for discriminating and counting erythroblasts

A method for classifying and counting leukocytes comprises the steps of: (1) adding to a hematological sample the following fluorescence-labeled antibodies labeled with fluorescent dyes which emit fluorescences distinguishable from each other; (a) a first fluorescence-labeled antibody (1st antibody) which bonds specifically to leukocytes, (b) a second fluorescence-labeled antibody (2nd antibody) which bonds to at least one kind of neutrophilic cells, and (c) a third fluorescence-labeled antibody (3rd antibody) which bonds to at least one kind of immature granulocytic cells, in order to stain leukocytic cells in the sample, and removing erythrocytes from the sample; (2) analyzing the resulting sample using a flow cytometer to measure at least one scattered light signal and three separate fluorescence signals; (3) defining a group of granulocytic cells on the basis of intensity of the scattered light and intensity of fluorescence from the 1st antibody; (4) defining neutrophilic cells in the defined group of granulocylic cells on the basis of the intensity of the fluorescence from the 1st antibody and intensity of fluorescence from the 2nd or 3rd antibody; (5) classifying the defined group of the neutrophilic cells into groups of neutrophilic cells different in degree of maturity on the basis of the intensity of the fluorescence from the 2nd antibody and the intensity of the fluorescence from the 3rd antibody, and counting the number of cells in each of the groups.

Method for classifying and counting leukocytes

A process for counting sperms (sperm cells) comprises the steps of: (1) mixing a specimen containing sperms with an aqueous solution containing a cationic surfactant in an amount effective in removing contaminants contained in the specimen to give a surfactant-treated specimen, (2) staining the sperms contained in the surfactant-treated specimen with a staining liquid containing a dye for staining nucleic acid to give a measuring specimen, (3) introducing the measuring specimen into a flow cell in a flow cytometer and irradiating the stained sperms in the measuring specimen with excitation light, (4) detecting a scattered light signal and a fluorescent light signal emitted from the sperms irradiated with the excitation light, (5) preparing a two-dimensional distribution involving two axes of a scattered light intensity and a fluorescent light intensity based on the scattered light signal and the fluorescent light signal, and (6) specifying a region of a sperm mass on the two-dimensional distribution and counting the number of sperm in the region of the sperm mass.

Process for counting sperms and reagent therefor

Aqueous blood-sample diluting reagent and method of its use for compelling a morphological change in a blood sample to yield an MCV value assayed at elapsed time after the sample is drawn to be consistent within a diagnostically acceptable range with the original, immediate post-drawing MCV value. Selection of a small amount of a predetermined surfactant added within a limited range of concentration, and of a salt for adjusting osmotic pressure of the sample is thereby determined. The blood sample is treated with an anti-coagulant agent immediately post-drawing, and for assay in a particle analyzer at post-drawing elapsed time is diluted with the reagent solution. The reagent has an osmotic pressure (&pgr;) of approximately 150-400 mOsm/kg and a pH of 6.0-8.5. The surfactant is present in a 0.0005% to 0.5% concentration and has a hydrophile-lipophile balance (HLB) of 10-20.

Diluting reagent and method compelling time-independent consistency in MCV assay

A reagent for measurement of leukocytes and hemoglobin concentration in the blood includes a cationic surfactant in an amount sufficient to lyse erythrocytes and denature hemoglobin, at least one of the following hemoglobin stabilizers:

Reagent for measurement of leukocytes and hemoglobin concentration in

A reagent for measuring immature leukocytes which comprises: (1) a polyoxyethylene-based nonionic surfactant having the general formula (I) in a sufficient amount capable of fixing cytoplasms and cell membranes of immature leukocytes:

Reagent for measuring immature leukocytes

Reagents for enabling leukocytes to be classified and counted lyse erythrocytes and act on leukocytes to stabilize them are disclosed. The reagents contain polyoxyethylene-based surfactants, and may contain hyperosmotic or hypoosmotic agents and solubilizing agents. One embodiment of the invention contains surfactants selected from the anionic and nonionic polyoxyethylene-based surfactants having 18-30 repeating oxyethylene units per molecule. A second embodiment of the invention contains nonionic polyoxyethylene surfactants only, having 20-100 repeating oxyethylene units per molecule, does not employ either saponin or quaternary ammonium compounds, and provides for adjustment of the pH of the resulting solutions. The invention allows leukocytes to be classified into as many as five types, each type to be quantified, and enables the detection of abnormal cells, when used in conjunction with particulate analysis flow cytometers employing the RF method and the DC method.

Method for classifying leukocytes and a reagent used therefor

A reagent for measuring the hemoglobin concentration and counting the number of leukocytes in a blood sample, which contains at least one of a quaternary ammonium salt or a pyridinium salt, having a concentration capable of hemolyzing erythrocytes in the blood and denaturing hemoglobin and at least one of cationic, nonionic or amphoteric surfactants or an oxidant capable of oxidizing heme in hemoglobin, the reagent being capable of dividing leukocytes into two or three fractions, for example, a fraction of lymphocytes, a fraction of monocytes, eosinophils and basophils, and a fraction of neutrophils.

Reagent for simultaneous determination of hemoglobin and leukocytes in

A reagent for and method of measuring leukocytes or hemoglobin or both in blood using a reagent which is water-soluble mixture containing:

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Hyogo, Japan

Yukio Hamaguchi