Abstract: The invention describes the preparation of preserved, non-infectious control cells. The control cells are prepared by modification, activation or otherwise changing selected normal leukocyte cells, tissue culture cells or zenogenic transplants in immunosuppressed animals to have characteristics of abnormal cells or by the addition of cells from an established cell line, which have been similarly modified or activated, to a normal leukocyte population. The preferred method of preserving the control cells is by lyophilization using an isotonic 10% trehalose solution.

Abstract: A rapid, simple method for preparing beads for calibrating flow cytometers which contain a known number of fluorophores per bead is presented. Briefly, the invention utilizes beads coated with a stable complex of a fluorophore and an enzyme. the enzymatic activity of a known number of beads gives an accurate measure of fluorophore density on those beads.

Abstract: The method and apparatus for detecting the presence of an organism in a sample of liquid involves collecting an extinction spectrum of liquid sample, deconvoluting the spectrum to obtain a particle size distribution for the sample, comparing the spectrum and the particle size distribution with, respectively, a control spectrum and a control particle size distribution for the organism, and determining from the comparisons whether the organism to be detected is present in the sample. The apparatus and method permit on-line real-time quantitative classification, identification, and viability assessment of an organism such as a harmful microorganism in a water supply line.

Abstract: An apparatus for producing blood component products. In one embodiment, a plurality of a predetermined type of blood component is harvested from a source of whole blood. At least two on-line yield determination techniques are utilized to determine the yield of the harvested blood components. One is a predetermined yield prediction technique and the second is a predetermined yield monitoring technique, each of which are individually calibrated in relation to a predetermined off-line yield determination technique. The predetermined yield prediction and monitoring techniques each provide the yield of the harvested blood components and each is then utilized to provide a determined yield. Consequently, when the harvested blood components are packaged the determined yield may be associated therewith, thereby providing a blood component product.

Abstract: A method of determining cell membrane strength is disclosed, using focused light of selected frequency and energy density. The method is also useful in diagnosing the presence of certain diseases in cells, by comparing the membrane strength of cells exposed to cell-characteristic altering drugs to the membrane strength of a control group of cells.

Abstract: A method for producing blood component products. In one embodiment, a plurality of a predetermined type of blood component is harvested from a source of whole blood. At least two on-line yield determination techniques are utilized to determine the yield of the harvested blood components. One is a predetermined yield prediction technique and the second is a predetermined yield monitoring technique, each of which are individually calibrated in relation to a predetermined off-line yield determination technique. The predetermined yield prediction and monitoring techniques each provide the yield of the harvested blood components and each is then utilized to provide a determined yield. Consequently, when the harvested blood components are packaged the determined yield may be associated therewith, thereby providing a blood component product.

Abstract: The invention is a rapid, continuous test for glycated hemoglobin using a non-equilibrium affinity binding method. Agarose beads derivatized with 3-aminophenylboronic acid specifically bind glycated hemoglobin. This solid phase is incorporated into a sample processor card, modified to mix and to separate the test solution from the solid phase prior to absorbance readings. Two absorbance readings are made on the test solution, one immediately after mixing the reagent/diluent with the specimen, and one after a significant amount of binding has occurred. A linear correlation between total glycated hemoglobin and hemoglobin A.sub.1c permits standardization and reporting of units equivalent to % hemoglobin A.sub.1c. Stable glycated hemoglobin solutions for use as standards in the assay, and a method for preparing the standards are also disclosed.

Abstract: An assay and sample mixture for the enumeration of fluorescently stained target components of a whole blood sample by an imaging instrument. The sample preparation method ensures that the amount of target components per unit of volume of the whole blood sample is preserved by elimination of certain non-quantitative preparation steps while producing an even hematocrit layer within a scan capillary. Typical target components include white blood cells that express certain surface antigens, such as CD-4 and CD-8 proteins. To inhibit aggregation of the red blood cells, a reagent is added to an aliquot of whole blood sample. The aliquot of whole blood is mixed and with a preselected amount of a fluorescent dye and ligand complex which tags the target components.

Abstract: The invention provides a method of quantitating reticulocytes with high sensitivity and high accuracy at low cost employing a fluorescent dye suitable for the staining of reticulocytes. A blood sample stained with a fluorescent dye composition containing TOTO-3, TO-PRO-3, POPO-3, PO-PRO-3, YOYO-1, YO-PRO-1, YOYO-3 or YO-PRO-3 as the fluorescent dye is exposed to light in the red-wave length region in the case of TOTO-3 or TO-PRO-3, light in the green region for POPO-3 or PO-PRO-3, light in the blue region for YOYO-1 or YO-PRO-1, or light in the orange region for YOYO-3 or YO-PRO-3 and the resultant fluorescent emission from the blood sample is measured to enumerate reticulocytes.

Abstract: A method and a device for the simultaneous and quantitative, flow cytometric analysis of nucleated red blood cells (NRBC) and white blood cells (WBC) in a whole blood sample.

Abstract: A method and apparatus are provided for selecting and analyzing a subpopulation of cells or cell objects for a certain parameter such as DNA using image analysis means. The cells are first stained with an alkaline phosphatase technique including a monoclonal antibody specific to a protein in at least one of the cell's cytoplasm or on a cell membrane, thereby marking any cells including the protein as to type. A second staining of the DNA in the nucleus is accomplished by a Feulgen technique that destroys the cell cytoplasm. After the staining and marking, the cells may then be gated using the image analysis means on the visual parameter such as colored DNA or colored antigen into a subpopulation that is to be measured. The selected cells may then be examined by digital image processing and measured for a parameter such as a true actual measurement of DNA in picograms. A quantitation of the measured parameter may be generated and provided.

Abstract: A reagent for analyzing leukocytes having at least one nonionic surfactant having an addition polymerization molar number of polyoxyethylene of 3 to 9, at least one cationic surfactant, and a buffer adjusting a pH value to 2.5 to 4.0, which can determine a cell size of and the morphological features of leukocytes contained in the blood sample.

Abstract: A microphotolysis and microphotoanalysis process applying precise quantities of focused light energy to a precise microscopic area of a very small cell sample to activate a chemical agent which permits measurement of cell fragility.

Abstract: A hematology control product comprising leukocyte analogs and an aqueous solution of a plasma substance for use in a blood counting instrument is described. The instrument employs a lytic reagent system for the lysable red blood cells in the control product. The plasma substance is in an amount effective to enable the differentiation of each of said leukocyte analogs relative to the physical attributes of the analogs, said physical attributes of the analog are similar to human leukocytes. Preferably, the plasma substance comprises cholesterol or its derivatives.A method for using the cell suspension media comprising an aqueous solution of a plasma substance is also described. The method provides a quality control to determine whether an instrument is operating within manufacturer's specifications.

Abstract: Aqueous solutions containing buffers and electrolytes are adjusted to levels required for calibration of both blood gas analyzers and ion selective electrolyte analyzers. A control material, composed of similar matrix, is adjusted to three levels of blood gas and electrolyte conditions. These quality control materials are used to monitor blood gas/electrolyte laboratory instrumentation.

Abstract: A multipurpose reagent system for rapid analysis of a whole blood sample allowing the determination of at least five classes of peripheral white blood cells, nucleated red blood cells, and lymphocyte immunophenotyping on automated hematology instrumentation. The multipurpose reagent system lyses red cells rapidly, while it concurrently fixes white cells and preserves surface antigens on lymphocytes. The multipurpose reagent system comprises from about 3 to 7 grams per liter of a non-quaternary ammonium salt, from about 0.04 to about 0.10 percent by volume of an aliphatic aldehyde with one to four carbons, from about 10 mM to about 20 mM of a non-phosphate buffer which is inert to the aliphatic aldehyde, and a sufficient amount of water to give a pH between 5.5 and 7.5 and an osmolality of between about 160 to about 310 mOsm per liter.

Abstract: A hematology control product comprising leukocyte analogs is described. The analogs comprise red blood cells which simulate at least two physical properties of human leukocytes. A method for making leukocyte analogs from blood cells having desired physical properties is also described. The process comprises expanding the cell volume, changing the hemoglobin content of the cell and fixing the cell. Generally, the monocyte and lymphocyte analogs leak hemoglobin from the cell while the eosinophil analog has the hemoglobin precipitated in the cell. A further method is described to use the control product to determine whether an automatic instrument is operating within manufacturer's specification.

Abstract: A lytic agent is described which when mixed with whole blood, lyses the red cells instantly, but does not affect the optical, light scattering properties of the white blood cells for at least 30 seconds. This lytic agent enables the performance of a five part leukocyte differential by flow cytometry and light scattering. In its preferred embodiment, the lytic agent incorporates 2-phenoxyethanol which combines the functions of leukoprotective and antimicrobial, TRITON X-100 (octylphenoxypolyethoxyethanol) a lytic and wetting agent and TRIS/HCl to provide pH buffering and to increase the solution's conductivity.

Abstract: The present invention is a method for blood analysis which provides a rapid treatment of a blood sample so that it can be analysed for counting and classification of leukocytes. Leukocytes are permeablized by treatment with a surfactant solution and labeled, preferably with a fluorescent dye. The method provides labeled leukocytes that can be counted and classified by optical means, including flow cytometry by analysis of a fluorescence signal.

Abstract: A method for determining the activation level of platelets and other types of blood cells that undergo activation by ELISA and other immunoassay techniques includes the step of reacting a sample containing the blood cells in a liquid phase with an excess quantity of an activation-specific primary antibody prior to allowing the cells in the sample to bind to a solid surface. This differs from typical ELISA procedures, wherein the cells are bound to the surface of a plastic test tube or microtiter plate prior to addition of the primary antibody. It has been discovered according to the invention that binding the cells to a surface prior to reaction with the primary antibody changes the activation level of the cells, making the assay less accurate.

Abstract: The present invention has multiple aspects that include an intermediate composition (i.e., a marked stock calibrator solution) for preparing diluted calibrator solutions that are marked in proportion to the amount of calibrator contained therein; a diluted (i.e., working) calibrator solution made therefrom, a series of diluted calibrator solutions that are visually colored in proportion to the concentration of calibrator therein; a method for performing a diagnostic assay that employs a series of marked calibrator solutions; a method for confirming that a stock solution of calibrator has been diluted correctly; and a method for indirectly confirming the concentration of calibrator in a working calibrator solution.

Abstract: A reagent composition which includes organic cationic dyes for staining the reticulocytes in a blood sample and buffer solutions for maintaining a pH of about 6 to about 9 is provided. The dyes may be the blue absorption dyes Oxazine 750 or New Methylene Blue. A zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes. The reagent composition and whole blood sample mixture are passed through the sensing region of a flow cytometer. The light scattered and absorbed by each cell is measured; the erythrocytes can be distinguished from reticulocytes and the volume, hemoglobin concentration and the hemoglobin content of each reticulocyte or erythrocyte, and the mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.

Abstract: A stable reticulocyte reference control which may be used with both manual and flow cytometric reticulocyte counting devices is provided. The reference control contains erythrocytes loaded with nucleic acids or polyanions capable of binding with a cationic dye. The loaded erythrocytes are suspended in an aqueous preserving medium. Methods of making and using the reticulocyte reference control are also provided.

Abstract: Aqueous solutions containing buffers and electrolytes are adjusted to levels required for calibration of both blood gas analyzers and ion selective electrolyte analyzers. A control material, composed of similar matrix, is adjusted to three levels of blood gas and electrolyte conditions. These quality control materials are used to monitor blood gas/electrolyte laboratory instrumentation.

Abstract: A cell fixative composition for fixing the internal components of a cell without disrupting the cell surface components is disclosed. The composition contains a compound for preserving intracellular morphology, a compound for fixing internal components of the cell, a compound which facilitates transportation of components across the cellular membrane, and a compound for increasing permeability of cell membranes. Methods for analyzing cells which have been fixed using the composition are also disclosed.

Abstract: A reagent composition which include an organic cationic dye for staining the reticulocytes in the blood sample and a buffer solution for maintaining a pH of about 6 to about 9 is provided. The dyes may be the red excitable fluorescent dye Oxazine 750, or the blue excitable fluorescent dyes Acridine Orange or derivatives of Acridine Orange. A zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes.

Abstract: The invention describes the preparation of preserved, non-infectious control cells. The control cells are prepared by modification, activation or otherwise changing selected normal leukocyte cells, tissue culture cells or zenogenic transplants in immunosuppressed animals to have characteristics of abnormal cells or by the addition of cells from an established cell line, which has been similarly modified or activated, to a normal leukocyte population. The preferred method of preserving the control cells is by lyophilization using an isotonic 10% trehalose solution.

Abstract: In a cell analyze apparatus, a light beam is irradiated onto cells (or particles like the cells) flowing through a flow cell so as to measure cell light information for each cell with respect to a plurality of parameters (for example, the forward scattered light intensity, the right angle scattered light intensity and the intensity of fluorescence by different dye). Based on a minimal point of a histogram associated with the cell light information with respect to one or more parameters, the cell population is subdivided into fractions. When the minimal point is missing in the histogram, the parameters above are converted by use of a predetermined conversion expression (for example, a coordinate conversion is effected on the parameters) such that a minimal point is detected from the histogram of cell light information related to the new parameters obtained by the conversion, thereby subdividing an objective cell population.

Abstract: A method for monitoring water filter performance and efficiency, by correlating particle size and distribution to turbidity measurement. The method includes the steps of installing and calibrating the filter system, and periodically testing the filter system efficiency. The calibration of the filter system includes the step of preparing a calibration "challenge" solution having a predetermined count of particles of known size which will serve as "surrogates" for the particles which are to be removed during normal filtration. The turbidity of the calibration challenge solution is measured, and correlated to the particle count information. Similarly, the effluent turbidity is measured and a particle size distribution count made to verify the percent removal specified.

Abstract: 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.

Abstract: A system for rapid microbead calibration of a flow cytometer including a suspension of quantitative fluorescent microbead standards and analytical software. The software is used to take information on the microbead suspension from a flow cytometer and analyze data, smooth curves, calculate new parameters and notify of expiration of the system.

Abstract: This invention primarily is directed to a hematology reference control solution, the three separate white cell control portions thereof consisting of three types of fixed red cells of determined size distribution for checking the operation of a particle analyzing instrument, including its predetermined lower and upper threshold settings for each class or subclass of leukocytes.For preparing a human granulocyte analogue, nurse shark erythrocytes are altered and fixed in a chilled solution to simulate in number, size and distribution the granulocytes in human whole blood.

Abstract: Methods for characterizing and distinguishing reticulocytes and mature red blood cells use reagent compositions which include an organic cationic dye for staining the reticulocytes in the blood sample and a buffer solution for maintaining a pH of about 6 to about 9. The dyes may be the red excitable fluorescent dye Oxazine 750, or the blue excitable fluorescent dyes Acridine Orange or derivatives of Acridine Orange.

Abstract: Methods for characterizing and distinguishing reticulocytes and mature red blood cells use reagent compositions which include organic cationic dyes for staining the reticulocytes in the blood sample and buffer solutions for maintaining a pH of about 6 to about 9. The dyes may be the blue absorption dyes Oxazine 750 or New Methylene Blue.

Abstract: The invention describes the preparation of non-infectious control cells from normal, non-infectious, non-disease altered blood by depletion or augmentation of one or more cell types found in normal blood to reflect a specific disease state. The control cells so produced are preserved and thereafter reconstituted for use in immunological assays.

Abstract: A stabilized, aqueous composition containing FK506. FK506 degrades rapidly in most aqueous matrices. The rate of degradation is decreased in the presence of unfixed blood cells or fragments of blood cells from human or animal sources. A number of matrices using blood components are possible. Blood can be used directly or the blood cells can be lysed. The blood is diluted with a solution of an alkali halide.

Abstract: A hematology control product comprising leukocyte analogs is described. The analogs comprise red blood cells which simulate at least two physical properties of human leukocytes. A method for making leukocyte analogs from blood cells having desired physical properties is also described. The process comprises expanding the cell volume, changing the hemoglobin content of the cell and fixing the cell. Generally, the monocyte and lymphocyte analogs leak hemoglobin from the cell while the eosinophil analog has the hemoglobin precipitated in the cell. A further method is described to use the control product to determine whether an automatic instrument is operating within manufacturer's specification.

Abstract: A method of setting up a flow cytometer using microbeads fluorescing at a plurality of wavelengths, control cells for each of said wavelengths, certified blank microbeads. The blank microbeads are run and the PMT voltages are adjusted so that the blank microbeads are on scale on the flow cytometer. The control cells are run and the photomultiplier tube voltages are adjusted so that the single population falls in selected channels. The fluorescent microbeads are run on the flow cytometer and the Target Channels for the flow cytometer are determined.

Abstract: A method which comprises using a specified dye taken up by erythrocytic nucleated cells so that their nuclei are stained and can be readily differentiated from other cells by measurement with a flow cytometer is disclosed. The method relies basically upon flow cytometry by two-step staining that uses a first fluid that is an acidic hypotensive fluorescent dye solution and a second fluid that is a solution that changes the osmorality and pH of the first fluid. The first fluid further contains a fluorescent dye that diffuses into erythroblasts to specifically stain their nuclei, and a group of erythroblasts is separated from other cell groups on the two-dimensional plot constructed with the flow cytometer, whereby the results of erythroblast differentiation are computed.

Abstract: A calibration standard for calibrating machines which count the somatic cells in a milk sample, comprises an aqueous dispersion of microbeads bearing a fluorescent dye, a suspending agent, and an electrolyte; the dye has an excitation wavelength below 580 nm and a fluorescence emission wavelength in the range of 550 to 660 nm, with the excitation wavelength being at least 10 nm below the emission wavelength; the microbeads are present in a predetermined number per unit volume of dispersion; typically the standard contains 1.times.10.sup.5 to 9.times.10.sup.5 beads/ml.

Abstract: Methods for blood sample preparation using reagent compositions which include a zwitterionic surfactant for sphering of cells to eliminate orientational noise. When the reagent composition including a for staining a subset of cells is reacted with an anticoagulated blood sample, and the reaction mixture is passed through the sensing region of a flow cytometer, the light scattered and absorbed by each cell is measured, the stained cells can be distinguished from the unstained cells, and when the cells are reticulocytes and mature erythrocytes, respectively, the volume and hemoglobin concentration of each reticulocyte or erythrocyte, and the hemoglobin content, mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.

Abstract: A procedure for establishing the presence of live housedust mites in a textile sheet-like article, in which the housedust mites are allowed to migrate onto the surface of the article to be tested or are caused to start such a migration, if appropriate by the action of heat, the housedust mites which have migrated being picked up at this point and subsequently identified.

Abstract: Reference controls comprised of aldehyde-fixed white blood cells stabilized red blood cells and simulated blood platelets exhibiting a white blood cell histogram profile that is substantially that of whole blood are obtained by the addition of a lipoprotein to the control and an antioxidant to inhibit lysis of stabilized red blood cells by said lipoprotein.

Abstract: Reference controls comprised of aldehyde-fixed white blood cells exhibiting a white blood cell histogram profile that is substantially that of whole blood are obtained by the additions of a lipoprotein to the control.

Abstract: A method and apparatus for automatically and rapidly, retrieving counting and/or analyzing at least one selected population of cells or formed bodies, such as a white blood cell population and at least one subset thereof of a whole blood sample or portion thereof. A volume of a biological medium containing the white blood cells is prepared and at least one reactant specific or preferential at least to some selected biological cells is introduced thereto and rapidly mixed for a short period of time. A multipart blood cell analysis is obtained with a single sensing parameter by depleting at least one WBC subset population. The percentage of a desired WBC population subset or the overlapping of WBC subset populations also can be obtained by subtracting one or more obscuring WBC subset populations.

Abstract: 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.

Abstract: A new method for separation of white cells in differential hematology instruments is described. In the method described, white cell shrinkage is controlled by the use of diazolidinyl urea and accelerated with alcohol thereby providing a reproducible method for easy manipulation of the white cells. In addition, it was found that adsorption of blood platelets and red blood cells in the instruments can be prevented by use of a surfactant such as the polyols.

Abstract: A reagent, satisfying the undermentioned conditions, is useful for counting the number of leukocytes and measuring the hemoglobin concentration in blood samples. The reagent is free of cyanides and it is able to maintain hemoglobin in a stable state for a prolonged time. The reagent contains at least one cationic surfactant selected from the group consisting of a quaternary ammonium salt of a concentration capable of hemolyzing erythrocytes in the blood and oxidizing hemoglobin and a composition of pyridinium salts; at least one cationic, nonionic or amphoteric surfactant; and at least one hemoglobin stabilizer.

Abstract: The present invention relates to a reagent and to a method of using the same for automatically determining at least one leukocyte sub-population from a total blood sample. Using this reagent, it is possible, in a single step, to lyse the erythrocytes through the action of saponin and SDS, as well as to ensure the differential staining of the leukocyte sub-populations, at the same time preserving the integrity and morphology of the cells, through the combined action of various components including chlorazol black, an alcohol, a surfactant and preserving agents. The reagent according to the invention can be applied to different types of automatic flow cytometry analyzers.

Abstract: A method and apparatus for automatically and rapidly, retrieving, counting and/or analyzing at least one selected population of cells or formed bodies, such as a white blood cell population and at least one subset thereof of a whole blood sample or portion thereof. A volume of a biological medium containing the white blood cells is prepared and at least one reactant specific or preferential at least to some selected biological cells is introduced thereto and rapidly mixed for a short period of time. The opacity and/or volume parameter of the cells can be modified and the mixture is then counted and analyzed in one or more steps to obtain the desired white blood cell population and subset analysis. The biological sample can be a whole blood sample and the reactant can include or be a lyse or a monoclonal antibody bound to microspheres, which will bind to specific ones of the cells or a combination of lyse and microspheres with antibody bound thereto.