Abstract: The invention provides methods and compositions for simultaneously detecting the activation state of a plurality of proteins in single cells using flow cytometry. The invention further provides methods and compositions of screening for bioactive agents capable of coordinately modulating the activity of a plurality of proteins in single cells. The methods and compositions can be used to determine the protein activation profile of a cell for predicting or diagnosing a disease state, and for monitoring treatment of a disease state.

Abstract: A method for enumerating white blood cells and nucleated red blood cells. The steps of the method are as follows: (a) providing a lysed sample of whole blood; (b) introducing the lysed sample to a light-scattering multi-angle depolarizing flow cytometer; (c) removing depolarizing interference, e.g., lipid droplets and other measured particles; (d) differentiating nucleated red blood cells and noise from white blood cells in the absence of depolarizing interference; (e) differentiating nucleated red blood cells from noise in the absence of depolarizing interference and white blood cells; and (f) differentiating possible platelet clumps.

Abstract: Methods for differentially identifying cells in an instrument employ compositions containing a combination of selected antibodies and fluorescent dyes having different cellular distribution patterns and specificities, as well as antibodies and fluorescent dyes characterized by overlapping emission spectra which form non-compensatable spectral patterns. When utilizing the compositions described herein consisting of fluorescent dyes and fluorochrome labeled antibodies with overlapping spectra that cannot be separated or distinguished based upon optical or electronic compensation means, a new fluorescent footprint is established. This new fluorescent footprint is a result of the overlapping spectra and the combined cellular staining patterns of the dyes and fluorochrome labeled antibodies chosen for the composition. The new fluorescent footprint results in histogram patterns that are useful for the identification of additional cell populations or subtypes in hematological disease.

Abstract: Methods and compositions comprising immunoassays for the detection of antigens and antibodies in a sample are described. In particular, the present invention provides assays that are useful for the rapid and simultaneous detection of multiple different antigens and antibodies. In preferred embodiments, the assays include fluorescent labels of multiple wavelengths or intensities, which are used to label the antigens and antibodies directly and to label beads coated with molecules specific for the antigen or antibody. The detection of a fluorescence shift indicates the presence or identity of the antigen or antibody in the sample.

Abstract: Disclosed is a single population of simulated leukocyte granules prepared from mammalian whole white blood cells, which is useful in calibrators for a hematology analyzer, a calibrator which includes the single population of simulated leukocyte granules. Disclosed is also a method of preparing the single population of simulated leukocyte granules. The method of preparing single population of simulated leukocyte granules includes the actions of: obtaining mammalian whole white blood cells; treating the mammalian whole white blood cells with a hemolytic agent and a leukocyte treating liquid which includes an isotonic saline solution. The isotonic saline solution may include a fixative, an oxidizer, and a carbohydrate compound; fixing the treated mammalian whole white blood cells with a fixative; and suspending the fixed cells in a preservation medium for long-term preservation of fixed cells.

Abstract: An assay technique for label-free, highly parallel, qualitative and quantitative detection of specific cell populations in a sample and for assessing cell functional status, cell-cell interactions and cellular responses to drugs, environmental toxins, bacteria, viruses and other factors that may affect cell function.

Abstract: A sample analyzer with a removable and/or disposable cartridge for use at the point of care of a patient such as in a doctor's office, in the home, or elsewhere in the field. By providing a removable and/or disposable cartridge with all of the needed reagents and/or fluids, the sample analyzer can be reliably used outside of the laboratory environment, with little or no specialized training.

Abstract: The invention relates to a reagent and a process for the identification and counting of biological cells in a sample. This reagent comprises a cell lysing agent selected from at least one detergent in a concentration capable of specifically lysing a given type of cells in the sample, and a stain capable of marking the intracellular nucleic acids of the remaining unlysed cells. Application in particular for the identification and counting of cells using an automated analysis system based on flow cytometry.

Abstract: A method for distinguishing erythrocytes in a biological specimen, includes preparing a sample liquid so as to cause damage to a cell membrane of yeast-like fungi without hemolyzing erythrocytes in a biological specimen and to stain the yeast-like fungi with a fluorescent dye; detecting a first information and a second information from a particle in the sample liquid, wherein the first information reflects a size of the particle and the second information reflects a degree of fluorescent staining of the particle; and distinguishing the erythrocytes from the yeast-like fungi based on the first information and second information detected. An apparatus, a reagent kit and a reagent for carrying out the method are also disclosed.

Abstract: A method useful for the enumeration of cell populations in a biological sample includes the steps of reacting in a single reaction mixture a sample, a first antibody labeled with a fluorochrome having a first emission spectrum and an additional antibody. The first antibody binds to an antigenic determinant differentially expressed on leukocytes and non-leukocytes. The additional antibody binds to an antigenic determinant differentially expressed on mature and immature granulocytes or myeloid cells, and is labeled either with the first fluorochrome or an additional fluorochrome having an emission spectrum distinguishable from the first emission spectrum. The reaction mixture can be mixed with a nucleic acid dye having an emission spectrum that overlaps with one of the first or additional emission spectra. The reaction mixture may be treated with a lytic system that differentially lyses non-nucleated red blood cells and conserves leukocytes.

Abstract: A reagent for analyzing immature leukocyte capable of analyzing accurately immature leukocyte such as myeloblast and immature granulocyte, and mature leukocyte, which reagent has broader allowable range of sample processing conditions than conventional ones, and a reagent kit are provided. The present invention relates to a reagent for analyzing immature leukocyte containing a surfactant for giving damage to cell membrane of red blood cell and mature leukocyte in the sample, a solubilizing agent for causing contraction to damaged blood cell, sugar, a dye for staining nucleic acid.

Abstract: Analyzers and methods of analysis are described for performing blood cell counting and immunoassay on a whole blood specimen in one measurement section. An assay sample is prepared by blending carrier particles sensitized with an antibody or an antigen against a substance to be immunoassayed and a fluorescent dye for staining blood cells with the whole blood specimen. Optical information is detected from a particle in the assay sample, and the blood cells are differentiated and counted based on the detected optical information. A rate of agglutination of the carrier particles is obtained based on the detected optical information, thereby enabling detection of the substance to be immunoassayed.

Abstract: A control and a cellular fraction for use therein and methods for making the same, including a first cellular component including a plurality of a first group of processed animal red blood cells other than human blood cells; a second cellular component including a plurality of a second group of processed animal red blood cells other than human blood cells; a third cellular component including a plurality of a third group of processed animal red blood cells other than human blood cells; wherein one or more of the first second and third cellular components include a nucleus; and the control simulates erythroblasts of a human blood sample on an automated blood analyzer that analyzes blood cells using impedance, optical measurements or both.

Abstract: A standard material that is used for judging an abnormal portion in a particle analyzer is described. The standard material comprises first standard particles to be fluorescence-stained by a fluorescence-staining treatment and second standard particles that have preliminarily contained a fluorescence dye. A method and an analyzer that can judge an abnormal portion in a particle analyzer by using such a standard material are also described.

Abstract: The invention provides compositions and methods for cell separation. These reagents and techniques specifically agglutinate cells via surface antigen recognition and can be used to recover rare cell types in high yield.

Abstract: The present invention provides a method for preparing a five-part differential white blood cell control and a white blood cell control prepared thereby. The white blood cell control obtained by the method of the present invention perfectly retains the light scattering properties of every type of white blood cells and has much higher stability than that of real white blood cells, and therefore can be used for the quality control of five-part differential hematology analyzers based on the principle of multi-angle light scattering.

Abstract: A method for identifying or monitoring SLE in an individual is provided. The method includes quantitating complement component C4d on the surfaces of platelets and comparing the amounts of C4d to reference levels of C4d on platelets of individuals without SLE and/or on platelets of the individual obtained at a different time. Kits for use in the above-described methods are provided along with computer readable media tangibly embodying executable instructions to perform the methods.

Abstract: The invention is related to methods of diagnosing inflammatory diseases or conditions by determining levels of components of the complement pathway on the surface of white blood cells.

Abstract: The present invention provides a reference solution for use in instruments that determine hematocrit levels in biological samples by measuring the resistance and/or conductivity of the biological samples. A reference solution according to the invention achieves conductivities representative of known hematocrit levels in blood, while maintaining tolerable levels of interference with the measurement of other analytes in the reference solution.

Abstract: The invention provides methods and compositions for simultaneously detecting the activation state of a plurality of proteins in single cells using flow cytometry. The invention further provides methods and compositions of screening for bioactive agents capable of coordinately modulating the activity of a plurality of proteins in single cells. The methods and compositions can be used to determine the protein activation profile of a cell for predicting or diagnosing a disease state, and for monitoring treatment of a disease state.

Abstract: The present invention relates to the qualitative and quantitative validation of marker indices, especially for medical diagnosis and particularly for the determination of the growth fraction in a sample with antibodies against the Ki-67 protein. Diagnostic kit for the quantification of a cell fraction labeled by a marker for in vitro diagnosis, characterized in that a pseudo-tissue is used for the intra- and inter-assay standardization of the marker index.

Abstract: The invention relates to a method and a device for detecting very small quantities of particles. The inventive method is based on a detection of antigen-antibody reaction products and provides a very high detection sensitivity all the way to the femtomolar or attomolar range.

Abstract: A method of single parameter and multiparameter characterizing of cells, particularly immunophenotyping of cells, is provided. The method preferably uses antibody coated microspheres which are adapted to bind to specific types of cells. One or more sets of coated microspheres are added simultaneously or sequentially to a suspension of cells and bind the cells they are adapted to bind. Cells may bind to one or more microspheres. The suspension is then filtered to trap bead-cell complexes. The complexes are preferably stained and then examined to characterize the cells, preferably the cells bound to the microspheres. A kit and apparatus for performing the method are also provided.

Abstract: Erythrocytes from a vertebrate animal other than a human that have been size reduced and treated with a fixing agent by conventional methods to resemble human platelets are further treated to achieve a size distribution that is detected in a consistent manner by different methodologies of blood cell counting. The further treatment includes heating to a denaturing temperature followed by a second treatment with a fixing agent. The resulting cells are useful as controls for calibrating and checking the accuracy of automated blood cell counting equipment.

Abstract: A rapid method for the quantitation of various live cell types is described. The method may include a variety of steps including: 1) suspending the cells in a detergent-like compound, 2) isolating the washed cells by centrifugation or filtration, 3) resuspending the cells in a solution that contains a preservative, a fluorescent dye and a compound such as dequalinium which can be taken up by the cells, 4) measuring the fluorescence increase over time of the cell-dye mixture with a simple fluorometer, and 5) measuring the native fluorescence of the cells. This new cell fluorescence method correlates with other methods of enumerating cells such as the standard plate count, the methylene blue method and the slide viability technique. The method is particularly useful in several applications such as: a) quantitating bacteria in milk, yogurt, cheese, meat and other foods, b) quantitating yeast cells in brewing, fermentation and bread making, c) quantitating mammalian cells in research, food and clinical settings.

Abstract: The invention is a method for analyzing immature reticulocytes for the presence of micronuclei. The method includes reticulocyte enrichment, fluorescent labeling, micronuclei staining, and analysis using single-laser flow cytometry. The invention also includes kits containing reagents to use in the method.

Abstract: A sample acquiring device for volumetric enumeration of white blood cells in a blood sample that includes a measurement cavity for receiving a blood sample. The measurement cavity has a predetermined fixed thickness. The sample acquiring device also has a reagent, which is arranged in a dried form on a surface defining the measurement cavity. The reagent has a hemolysing agent for lysing red blood cells in the sample and a staining agent for selectively staining white blood cells in the sample. Also, a system with the sample acquiring device and a measurement apparatus. The measurement apparatus has a sample acquiring device holder, a light source, and an imaging system for acquiring a digital image of a magnification of the sample The measurement apparatus also has an image analyzer arranged to analyze the acquired digital image for determining the number of white blood cells in the blood sample.

Abstract: A method for detecting aberrant phenotypes expressed by neoplastic cells includes the steps of: 1) staining one or more normal/reactive samples and one neoplastic sample with multiple combinations of monoclonal antibodies, 2) measuring fluorescence emissions associated to the stained cells, 3) storing two independent list mode data files of information on light scatter and fluorescence characteristics of each cell, 4) creating new data files by mixing list mode data from the data file containing information the neoplastic sample into the data file containing information on the normal samples, 5) defining corresponding to normal cells and areas corresponding to empty spaces in normal/reactive samples that may be occupied by tumor cells in neoplastic samples, 6) identifying events corresponding to neoplastic cells and events corresponding to normal cells coexisting in a multidimensional space, and 7) establishing the most relevant phenotypic aberrations displayed by the neoplastic cells as compared to their nor

Abstract: The present invention provides methods and systems for performing in vivo flow cytometry. In one embodiments, selected circulating cells of interest of a subject are labeled with fluorescent probe molecules. The labeled cells are irradiated in vivo so as to excite the fluorescent probes, and the radiation emitted by the excited probes is detected, preferably confocally. The detected radiation is then analyzed to derive desired information, such as relative cell count, of the cells of interest.

Abstract: A method and a system for monitoring the functional capability of a particle detector connected downstream, in the flow direction, of a particle filter, in which particles occurring upon the regeneration of the particle filter, in particular ions, are detected by the particle detector and the resultant measurement finding are compared with an expected finding. The measurement and evaluation are effected in a control and evaluation unit. The invention enables monitoring of the functional capability of a particle detector that does not interrupt normal operation and thus increases the reliability of the entire system.

Abstract: The invention relates to measuring of the adhesion of platelets ex vivo. In this method, monodisperse polymer particles are added to blood, and the number of platelets in the blood are measured before and after shaking of the sample. The level of adhesion of the platelets is determined by comparing the number of platelets prior to shaking with the number of platelets after shaking. Methods of establishing an adhesion index and diagnosis of a patient are also provided.

Abstract: Methods of prevention and correction of white blood cell interferences to the measurements of mean cell volume (MCV), red blood cell concentration (RBC) and hematocrit (Hct) of a blood sample are disclosed. The interference to MCV can be prevented by identifying a valley between the red blood cell and white blood cell modes on the red blood cell distribution histogram, defining a red blood cell region using the valley and calculating MCV within the defined region. Alternatively, a curve fit of the white blood cell population on the red blood cell distribution histogram can be used to exclude the white blood cells. RBC can be corrected by subtracting the white blood cell concentration obtained from the analysis of a second aliquot sample, when the white blood cell concentration (WBC) exceeds a predetermined criterion. Hct of the blood sample can be calculated using the obtained MCV and RBC.

Abstract: Red blood cells from a vertebrate are treated to make them effective components of a hematology control, allowing the control to be used for detecting all blood cell components, including white blood cells and platelets. The treatment includes the use of a fixative under limited conditions of concentration and exposure time, and the resulting red blood cells are stable but lysable in a hematology instrument and have a reduced tendency to form particulates.

Abstract: The invention provides compositions and methods for cell separation. These reagents and techniques specifically agglutinate cells via surface antigen recognition and can be used to recover even rare cell types in high yield.

Abstract: The present invention discloses a thin reagent containing film sensor for detecting and measuring free chlorine in water, where components of the film sensor are a polymeric substrate that contains reactive material, an organic polyhydroxy compound, a reagent that creates an associated polymeric matrix, and an indicator; and a method for making the same. The film sensor can be formed to fit a specific dimension or shape. The film sensor swells or dissolves when exposed to aqueous solutions so that said reagent is released so that it can react with free chlorine, or the film sensor swells when exposed to aqueous solutions so that the aqueous solution diffuses into the film sensor and reacts with said reagent contained within the swollen film sensor.

Abstract: A method for classifying leukocytes in animal blood is described. In the method, a measurement sample is prepared by mixing a canine or feline blood sample with a lysing reagent. Erythrocytes are lysed and leukocytes are shrunk in the measurement sample. The data correlated with the size of leukocytes in the measurement sample are measured. The leukocytes, on the basis of the measured data, are classified into a first group containing lymphocytes, a second group containing neutrophils and monocytes and a third group containing eosinophils.

Abstract: The present invention provides a method for preparing a five-part differential white blood cell control and a white blood cell control prepared thereby. The white blood cell control obtained by the method of the present invention perfectly retains the light scattering properties of every type of white blood cells and has much higher stability than that of real white blood cells, and therefore can be used for the quality control of five-part differential hematology analyzers based on the principle of multi-angle light scattering.

Abstract: The present invention discloses a single population of simulated leukocyte granules prepared from mammalian whole white blood cells, which is useful in calibrators for a hematology analyzer, a calibrator comprising the single population of simulated leukocyte granules, and a method of preparing the single population of simulated leukocyte granules. The method of preparing single population of simulated leukocyte granules according to present invention comprises the steps of: obtaining mammalian whole white blood cells; treating the mammalian whole white blood cells under the action of a hemolytic agent in the presence of a leukocyte treating liquid, i.e., an isotonic saline solution comprising a fixative, a oxidizer and a carbohydrate; fixing the treated mammalian whole white blood cells with a fixative; and suspending the fixed cells in a preservation medium suitable for long-term preservation of fixed cells.

Abstract: A lytic reagent composition and methods for differential analysis of leukocytes are disclosed. In embodiments, the analysis may utilize optical measurements in flow cytometry based hematology analyzers. The reagent system includes an anionic surfactant in a hypotonic solution, an inorganic buffer to maintain the pH in a range from about 6 to about 10, and optionally a leukocyte stabilizer. The reagent system is used to lyse red blood cells and stabilize the leukocytes to enable multi-part differentiation of leukocytes in a near physiologic pH environment on a flow cytometry based hematology analyzer using axial light loss, light scatter intensity, high-numerical aperture side scatter, and time-of-flight measurements.

Abstract: A self-calibrated, magnetic binding assay (e.g., sandwich, competitive, etc.) for detecting the presence or quantity of an analyte residing in a test sample is provided. The magnetic binding assay includes detection probes capable of generating a detection signal (e.g., fluorescent non-magnetic particles) and calibration probes capable of generating calibration signal (e.g., fluorescent magnetic particles). The amount of the analyte within the test sample is proportional to the intensity of the detection signal calibrated by the intensity of the calibration signal.

Abstract: A reagent for classifying leucocytes, including: a) at least one of surfactant selected from a cationic surfactant, an amphoteric surfactant, and the combination thereof; b) an organic compound having phenyl or heterocyclic group, wherein when the surfactant is a cationic surfactant, the organic compound is an organic compound having an anionic group; when the surfactant is an amphoteric surfactant or a combination of amphotheric and cationic surfactant, the organic compound is an organic compound having an anionic or cationic group; c) a buffer for adjusting pH.

Abstract: The present invention relates to a reagent for classifying leukocytes including (a) at least one surfactant capable of lysing erythrocytes and partly damaging the cell membrane of leukocytes, (b) at least one organic compound bearing an anionic group capable of binding to the cationic component present in the leukocytes to provide morphological differences between the leukocytes, and (c) a buffer for adjusting pH to 2˜8. Also disclosed is a method for classifying leukocytes into four groups.

Abstract: Methods and devices are described for monitoring a therapeutic effect in a red blood cell-related disease by detecting the size of a reticulocyte using a full-automatic blood cell counting device.

Abstract: A method of using a reference control for measurement of immature granulocytes on a blood analyzer is disclosed. The method includes analyzing a reference control that contains an immature granulocyte component made of processed red blood cells for simulating immature granulocytes and a mature white blood cell component for simulating white blood cells of a blood sample on a blood analyzer adapted for measurement of immature granulocytes, and reporting the immature granulocyte component and the mature white blood cell component of the reference control.

Abstract: The present invention provides an approach for the simultaneous determination of the activation states of a plurality of proteins in single cells. This approach permits the rapid detection of heterogeneity in a complex cell population based on activation states, and the identification of cellular subsets that exhibit correlated changes in activation within the cell population. Moreover, this approach allows the correlation of cellular activities or properties. In addition, the use of potentiators of cellular activation allows for characterization of such pathways and cell populations.

Abstract: Methods for diagnosing and monitoring systemic lupus erythematosus (SLE) or scleroderma by determining, in a blood sample from the individual being diagnosed or monitored, complement component C4d deposited on surfaces of red blood cells in the sample, and optionally also determining complement receptor CR1 deposited on the red blood cell surfaces. For diagnosis this is compared with the quantity of C4d (and optionally CR1) present on red blood cells of normal individuals. For monitoring it is compared with a value in a sample or samples previously obtained from the individual patient. The comparison may be made with individual values for C4d and CR1 and/or with a ratio of the two found in normal individuals.

Abstract: An immunoassay comprises the steps of: (a) mixing a whole blood sample with sensitized insoluble carrier particles smaller than erythrocytes to cause an immune agglutination reaction; (b) introducing the resulting immune agglutination reaction mixture including agglutinated particles and unagglutinated particles to a flow cell, irradiating the particles passing through the flow cell with laser light, and detecting scattered lights generated thereby; (c) setting a threshold value for distinguishing unagglutinated particles from agglutinated particles and a threshold value for distinguishing the agglutinated particles from blood cells with regard to intensity of the scattered light; and (d) distinguishing and counting the unagglutinated particles, the agglutinated particles and the blood cells from the scattered lights detected in the step (b), in reference to the threshold values set in the step (c).

Abstract: The present invention relates to a hematology instrument system that discriminates platelets from red blood cells, debris and other particles within a blood sample. The instrument system uses an optical trigger and collects data at optical sensor locations relative to a flow cell-illuminating laser beam's optical axis. An axial sensor measures axial light loss due to a particle in the flow cell's illumination aperture. In addition, the instrument system can include an RF unit generates electrical parameters, such as DC and RF parameters, within the flow cell. Blood specimens can be prepared with and without a sphering agent.

Abstract: Methods and kits for preparing and adjusting pre-defined master calibration curves of the type used for quantifying analyte polynucleotides by real-time nucleic acid amplification. Particularly disclosed are methods of preparing one or more master calibration curves on one instrument, and then using those master calibration curves on a different instrument.

Abstract: The invention provides methods and compositions for simultaneously detecting the activation state of a plurality of proteins in single cells using flow cytometry. The invention further provides methods and compositions of screening for bioactive agents capable of coordinately modulating the activity of a plurality of proteins in single cells. The methods and compositions can be used to determine the protein activation profile of a cell for predicting or diagnosing a disease state, and for monitoring treatment of a disease state.