The invention consists of compositions and methods using low density lipoprotein to reduce agglutination and improve the health of sex-sorted sperm cells.
The invention includes a method of screening a biological sample for contamination by sequencing a first biological sample at a first depth to generate a first nucleotide sequence; determining map coordinates for one or more homozygous loci in the first nucleotide sequence; sequencing a second biological sample at a second depth to generate a second nucleotide sequence, wherein the second depth is lower than the first depth; determining loci in the second nucleotide sequence at the determined map coordinates; and comparing the one or more homozygous loci in the first nucleotide sequence with the determined loci in the second nucleotide sequence.
C12Q 1/689 - Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
G16B 20/20 - Allele or variant detection, e.g. single nucleotide polymorphism [SNP] detection
G16B 30/00 - ICT specially adapted for sequence analysis involving nucleotides or amino acids
The invention includes a method of evaluating a bovine embryo by fertilizing an egg obtained from a first bovine heterozygote of a recessive lethal haplotype with sperm cells obtained from a second bovine heterozygote of the recessive lethal haplotype; producing the embryo from the fertilized egg, wherein the embryo is homozygous for the lethal haplotype; establishing a cell culture from the embryo; collecting a plurality of cultured cells; and obtaining omics data, comprising one or more features, from the plurality of cultured cells.
C12N 15/00 - Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
A discrimination system that forms a fluid column and interrogates objects within the fluid column with an excitation source. An optical arrangement collects output electromagnetic radiation emanating from the excited objects disposed within the fluid column and directs the output electromagnetic radiation to a detector. An analyzer reduces the positional dependency of the detected intensity by normalizing the value based on the position of each object.
C12M 1/34 - Measuring or testing with condition measuring or sensing means, e.g. colony counters
C12Q 1/04 - Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
The invention provides serialized artificial insemination straws, as well as systems and methods for authentication of the serialized artificial insemination straws.
The invention encompasses methods for reducing the proportion of sperm cells with abnormal morphology, and unviable sperm cells, in a sperm cell population.
An optical arrangement receives output light emanating from an object disposed within a fluid column that crosses an optical refraction boundary of the fluid column between the object and the optical arrangement. The optical arrangement modifies the output light such that the modified output light has an intensity that is more uniform than the output light.
The invention encompasses a rapid and safe preparation method of sperm nuclei, improved sperm nuclei and a method of using the improved sperm nuclei to calibrate a flow cytometer with higher accuracy.
The invention consists of methods and compositions for detecting the presence or absence of a DNA aberration by analyzing fluorescence emission characteristics in sperm cells or sperm nuclei, which generally consists of entraining sperm cells or sperm nuclei stained with a DNA selective dye in sheath fluid; exposing the entrained sperm cells or sperm nuclei to electromagnetic radiation; determining a forward fluorescence characteristic and a side fluorescence characteristic of individual events associated with the exposed sperm cells or sperm nuclei; and gating the individual events based on the forward fluorescence characteristic and the side fluorescence characteristic with a criterion.
Within the field of animal breeding, there is a need to rapidly develop in-bred animal populations and subsequently generate hybrids from those in-bred populations. The present invention does this by differentiating sperm and egg cells from stem cells or induced pluripotent stems cells obtained from embryos, thereby dramatically reducing the time needed to produce an in-bred population and subsequent hybrids.
This disclosure pertains to analytical instruments and related methods incorporating beam shaping optics for differentiating very bright and closely related signals over a wide range of operating conditions with an improved and uniform performance.
The invention encompasses a method of estimating a distribution or a standard deviation of breeding values or predicted transmitting abilities of gametes from an individual using population-wide estimates of linkage disequilibrium for one or more genetic markers or genes, so as to more efficiently produce offspring having breeding values or predicted transmitting abilities that equal or exceed a desired breeding value or predicted transmitting ability.
Embodiments of the present invention relate generally to processes, systems, and compositions useful in manipulating a ratio of viable X chromosome bearing sperm to viable Y chromosome bearing sperm in at least one sperm population and useful for preserving the resulting manipulated sperm population. In some embodiments a cryoprotectant may be incorporated into various medias used in manipulating the sperm sample, such as in a staining media, a sheath fluid, and a collection media.
Embodiments of the present invention relate generally to processes, systems, and compositions useful in manipulating a ratio of viable X chromosome bearing sperm to viable Y chromosome bearing sperm in at least one sperm population and useful for preserving the resulting manipulated sperm population. In some embodiments a cryoprotectant may be incorporated into various medias used in manipulating the sperm sample, such as in a staining media, a sheath fluid, and a collection media.
The invention encompasses methods for increasing genetic progress in livestock, and for genetic dissemination, including the use of amniocentesis to obtain fetal amniocytes for use in genomic evaluation and cloning.
An animal lift consisting of a lift mechanism, a vertical mast housing at least a portion of the lift mechanism, a horizontal shaft rotatably coupled to the lift mechanism about a longitudinal axis of rotation of the shaft, and a cradle coupled to the shaft, the cradle constructed of a horizontal support member having a surface for engaging the sternum or underbelly of an animal along the animal's longitudinal axis from head to tail and a pair of vertical side walls.
A01K 1/08 - Arrangements for simultaneously releasing several animals
A01K 13/00 - Devices for grooming or caring of animals, e.g. curry-combs; Fetlock rings; Tail-holders; Devices for preventing crib-biting; Washing devices; Protection against weather conditions or insects
A01K 15/00 - Devices for taming animals, e.g. nose-rings or hobbles; Devices for overturning animals in general; Training or exercising equipment; Covering boxes
A01K 15/04 - Devices for impeding movement; Devices for impeding passage through fencing
A22B 1/00 - Apparatus for fettering animals to be slaughtered
A22B 5/00 - Accessories for use during or after slaughtering
A22B 5/06 - Slaughtering stands or spreaders for cattle
The invention encompasses methods for increasing genetic progress in livestock, and for genetic dissemination, including the use of amniocentesis to obtain fetal amniocytes for use in genomic evaluation and cloning.
The present invention relates to compositions comprising low sugar media, the methods of using these compositions to reduce trauma and stress on processed animal sperm, the resulting sperm and embryo products, and the methods of use of these products to increase the quality, quantity and viability of progeny and improved rates of births in animals.
The present invention relates to compositions comprising low sugar media, the methods of using these compositions to reduce trauma and stress on processed animal sperm, the resulting sperm and embryo products, and the methods of use of these products to increase the quality, quantity and viability of progeny and improved rates of births in animals.
Embodiments of the claimed invention relate to sex sorted sperm having an improved dose response which may be produced by the steps of extending a sperm sample with a buffering holding media and adjusting the concentration of the extended sperm sample to a target concentration range. The extended sperm may then be stained with a DNA selective dye and sex sorted with a flow cytometer into a catch fluid. During the extending and sorting of the sperm sample, the pH may be maintained at a target pH. Additionally, at least one of the buffering holding media, the DNA selective dye and the catch fluid may be supplemented with at least one antioxidant.
A61D 19/00 - Instruments or methods for reproduction or fertilisation
A61P 15/08 - Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
23.
APPARATUS AND METHODS FOR HIGH THROUGHPUT SPERM SORTING
This disclosure relates to a system, device, and method for sorting sperm cells in a microfluidic chip. In particular, various features are incorporated into microfluidic chips and into sorting systems for aligning and orienting sperm in flow channels, as well as, for determining sperm orientation and measuring relative DNA content.
The invention relates to methods of increasing genetic merit of swine using sex sorted semen by establishing a plurality of mating subtypes for a line of swine, and determining a percentage of progeny that are male for each of the mating subtypes, or a percentage of progeny that are female for each of the mating subtypes, that would result, relative to a control, in an increase in genetic merit in the line.
A container suitable for storing biological material which is laser etched with a two-dimensional bar code and methods for producing the same. The container can be in the form of a straw having a thickness between about 0.1 mm and about 0.3 mm or can be in the form of another container that holds multiple straws. The laser etched mark can be in the form of a two-dimensional bar code may be located on an exterior surface of the container, and when the container is a straw, it remains unwarped and impermeable to fluids.
Methods for processing and sorting sperm are disclosed. Portions of sperm sorting or staining processes may include standardizing sperm samples by adjusting the concentration of the sperm sample to a predetermined concentration and adjusting the pH of the sample to a predetermined value. Sperm may also be stained in a single staining buffer having a DNA selective dye and a quenching dye.
This disclosure relates to cell sorting methods, and particularly cell sorting methods that improve the efficiency or productivity of sorting in a particle sorting instrument utilizing a measured parameter of sorting efficiency. In one embodiment, minimum productivity and minimum purity may be established and maintained while attempting to maximize the sorting efficiency. While in another embodiment, a minimum sorting efficiency and a minimum purity may be established and maintained while attempting to maximize the productivity of a sort.
A method of manufacturing a nozzle assembly may include the step of over molding a nozzle housing, or a portion of a nozzle housing, onto at least one nozzle component, such as an injection tube. Nozzle assemblies and flow cytometers incorporating nozzle assemblies may include any combination of straight smooth injection tubes, improved features for securing a nozzle assembly, improved features for debubbling a nozzle assembly, and aggressive orienting geometries. A method of sorting cells may include the step of magnetically coupling a nozzle assembly with a flow cytometer.
B29C 45/14 - Injection moulding, i.e. forcing the required volume of moulding material through a nozzle into a closed mould; Apparatus therefor incorporating preformed parts or layers, e.g. injection moulding around inserts or for coating articles
A nozzle tip formed from a cylindrical body defining a longitudinal axis and a frustoconical body adjoining the cylindrical body on the longitudinal axis. The cylindrical body may be in fluid communication with the frustoconical body. The frustoconical body may end in a flat surface with a nozzle exit orifice which is transverse to the longitudinal axis. There may be a cutout at the edge of the frustoconical body and the flat surface. The flow cytometer system may also include a source of electromagnetic radiation for producing a beam incident upon the fluid stream and the particles and a detector for detecting light emitted or reflected from the particles within the fluid stream in response to the beam.
A method of manufacturing a nozzle assembly may include the step of over molding a nozzle housing, or a portion of a nozzle housing, onto at least one nozzle component, such as an injection tube. Nozzle assemblies and flow cytometers incorporating nozzle assemblies may include any combination of straight smooth injection tubes, improved features for securing a nozzle assembly, improved features for debubbilng a nozzle assembly, and aggressive orienting geometries. A method of sorting cells may include the step of magnetically coupling a nozzle assembly with a flow cytometer.
A method for magnetic cellular manipulation may include contacting a composition with a biological sample to form a mixture. The composition may include a plurality of particles. Each particle in the plurality of particles may include a magnetic substrate. The magnetic substrate may be characterized by a magnetic susceptibility greater than zero. The composition may also include a chargeable silicon-containing compound. The chargeable silicon-containing compound may coat at least a portion of the magnetic substrate. The biological sample may include cells and/or cellular structures. The method may also include applying a magnetic field to the mixture to manipulate the composition.
C12N 1/00 - Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
C12Q 1/04 - Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
G01N 33/48 - Biological material, e.g. blood, urine; Haemocytometers
34.
METHODS FOR INCREASING GENETIC PROGRESS IN A LINE OR BREED OF SWINE USING SEX-SELECTED SPERM CELLS
The invention relates to methods of increasing the genetic progress of a line, breed or herd of swine through the use of sex-selected sperm cells in artificial insemination techniques. The invention also encompasses methods of artificially inseminating a swine via deep intrauterine catheter or via a laparoscopic procedure, which allow the use of reduced doses of sex-selected sperm cells.
A01K 67/00 - Rearing or breeding animals, not otherwise provided for; New breeds of animals
A01K 21/00 - Devices for assisting or preventing mating
A61D 19/02 - Instruments or methods for reproduction or fertilisation for artificial insemination
A61K 31/57 - Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
A61P 15/00 - Drugs for genital or sexual disorders; Contraceptives
A61P 15/08 - Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
35.
COMPOSITIONS AND METHODS FOR IMPROVING THE QUALITY OF PROCESSED SPERM
The present invention relates to compositions and methods for the handling of processed sperm including samples that are freshly collected, those transported as fresh samples, samples that are frozen and thawed, those sorted into one or more subpopulations, and those that are otherwise processed or handled that impose trauma on the cell. Trauma can reduce the motility, fertility, viability and overall integrity of the sperm and reduce the ability to fertilize, produce an embryo and a healthy offspring. The present invention relates to novel compounds that can be added to the sperm cell sample to reduce the traumatic effects of physical stress during mild as well as extensive sperm cell processing, methods of using the compounds in standard sperm processing procedures, the end products made from these methods including sperm and embryos, as well as methods of using those end products in assisted reproductive biology techniques in animals.
Generally, polymeric members and laser marking methods for producing visible marks on polymeric members, such as on thin and/or curved surfaces. The laser marking methods can include methods of laser marking straws with the step of matching laser source properties to the properties of straws being marked or with the step of laser marking straws having photochromic dyes.
A method of liquid nitrogen surface vitrification requiring an embryo washed in a rinsing medium, then incubated in a base medium and incubated in a hold medium before being washed in a vitrification medium and produced into a vitrification droplet (270). For forming the droplet, vitrification medium (210), an intermediary fluid such as air, followed by vitrification medium containing at least one embryo (250) are aspirated into the channel. The vitrification droplet consequently can contain an air bubble (220). The vitrification droplet can be produced from an instrument with a channel and dropped directly into liquid phase nitrogen producing a vitrified droplet. The vitrified droplet can be stored in cryo-vessels, and warmed for revitalization of biological function of vitrified biological cell mass or tissues, such as oocytes and/or embryos.
Generally, a computer implemented remote monitoring system which generates a viewable reduced byte data representation for each one of a plurality of analyzed instrument signals. Specifically, a flow cytometer remote monitoring system which generates a viewable reduced byte data representation for each one of a plurality analyzed flow cytometer signals.
The present invention relates to a method of remotely monitoring a flow cytometer coupled to a first computer, where the first computer produces a plurality of images as time bound data representation files based on signals generated in the flow cytometer and which are representative of the functional condition of the flow cytometer, the first computer being in communication with a second computer that converts the plurality of time bound data representation files into reduced byte data representation files with an image processor for storage.
Processes for selectively enriching a population of viable sperm cells with respect to a characteristic without physically sorting the cells are disclosed. The cells contained in such an enriched population benefit from the advantage of not being subjected to a sorting process. Processes of inseminating a female mammal and processes of forming a sperm dispersion utilizing the processes of selectively enriching a population of viable sperm cells are also disclosed.
A61B 17/425 - Gynaecological or obstetrical instruments or methods for reproduction or fertilisation
A61D 19/00 - Instruments or methods for reproduction or fertilisation
C12Q 1/02 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
41.
USE OF A COMPOSITION WHICH REGULATES OXIDATION/REDUCTION REACTIONS INTRACELLULARLY AND/OR EXTRACELLULARLY IN A STAINING OR SORTING PROCESS OF SPERMATOZOA
Staining mixtures comprising viable spermatozoa, a composition which regulates oxidation/reduction reactions intracellularly or extracellularly, and a DNA selective dye are disclosed. The cells contained in such suspensions tend to have a greater capacity for enduring the various process steps typically associated with the sorting of sperm cells into gender enriched populations, thereby resulting in post-sort compositions with an increased number of viable or motile sperm. Processes for staining sperm cells comprising the formation of a staining mixture are also disclosed.
C12Q 1/04 - Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
G01N 33/52 - Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper
The present invention provides a sperm dispersion comprising immotile spermatozoa, a composition that induces sperm immotility, and a cryopreservative.
Processes of storing sorted and unsorted spermatozoa, in the form of a sperm dispersion, having reduced motility relative to endogenous ejaculated sperm are disclosed. The immotile spermatozoa of the dispersion tend to have a greater capacity for enduring the rigors associated with storage, transportation, and fertilization procedures. Processes for providing such sperm dispersions, for inseminating a female mammal with such sperm dispersions, as well as compositions and combinations comprising the sperm dispersions, are also disclosed.
Sperm cell suspensions comprising a motility inhibitor are disclosed. The cells contained in such suspensions tend to have a greater capacity for enduring the various process steps typically associated with the sorting of sperm cells into gender enriched populations, thereby resulting in post-sort compositions with an increased number of viable or motile sperm. Processes for forming such cell suspensions, as well as processes for staining sperm cells, are also disclosed.
Apparatus for sorting stained particles in a fluid stream according to one or more characteristics of the particles, said system comprising: a. a fluid delivery system for delivering a fluid stream containing the particles to a first location; b. an electromagnetic radiation source for delivering a beam of electromagnetic radiation to the first location for exciting particles in the fluid stream to produce fluorescence emissions; c. an epi-illumination optics system including a focusing lens, the optics system being operable to direct the beam of electromagnetic radiation through the focusing lens in a forward direction along a beam axis intersecting particles in the fluid stream at said first location so that said particles pass through the beam, resulting in the fluorescence emissions from the particles being directed along said beam axis in a rearward direction; d. a photodetector for the detecting fluorescence emissions from the excited particles; and e. a processor in communication with the photodetector to classify the particles according to the one or more characteristics based upon the fluorescence emissions of the particles and to sort the particles according to the classification of particles.
A system for sorting a mixture of stained particles, including stained particles having a characteristic A and stained particles having a characteristic B, into at least one population, the system comprising: a. a fluid delivery system for delivering a fluid containing the stained particles in a flow path; b. an electromagnetic radiation source for exciting fluorescence emissions from the stained particles having characteristic A and the stained particles having characteristic B in the flow path; c. a photodetector for detecting the fluorescence emissions from the stained particles; d. a processor in communication with the photodetector for classifying the stained particles in the flow path according to their fluorescence emissions as either stained particles having characteristic A or stained particles having characteristic B; e. a control in communication with the detector for selecting stained particles to ablate; and f. a photo damaging laser in communication with the control for ablating selected particles in the flow path.
A flow cytometry apparatus nozzle to sort sperm cells in a fluid stream by chromosome content comprising: a. a nozzle body having a fluid stream entry portion and a fluid stream exit portion and an interior surface separating both portions and along which a fluid stream is adapted to flow; b. the interior surface of the nozzle including a first, a second and a third axially tapered region for progressively accelerating the speed of a fluid stream in a downstream direction toward the exit portion, at least two of the regions having generally elliptical cross sectional shapes oriented in different directions for applying torsional forces to a fluid stream passing therethrough, tending to bring sperm cells in the stream into a desired orientation.
Apparatus and methods for analyzing particles, including apparatus and methods for a sperm sorting process including: collecting sperm from an animal (30); selecting staining conditions (47A); staining the sperm with DNA selective fluorescent dye (48); sorting the sperm cells according to sex chromosome content (55); and cryopreserving a population of sorted sperm (61) until used for artificial insemination. One embodiment includes apparatus (1001) and methods for using a plurality of flow cytometry units (9) sharing an integrated platform to sort sperm cells. In one embodiment, flow cytometric sorting includes use of the following apparatus and methods: an orienting nozzle having a baffle (101); an epi-illumination optics system (109); slit scanning of localized DNA regions within cell nuclei (225); digital signal processing, including synchronous sampling of analog output signals (701), pulse waveform (497) feature extraction of an approximation of a first order derivative of a pulse waveform (497) at a point of the pulse, any of various sort strategies; and an automated sort calibration system (4201). In one embodiment, digital signal processing includes sampling analog output signals (701) at times relative to emission of pulses from an illumination laser. Other embodiments are substantially different from the foregoing, including embodiments directed to individual steps or systems that can be used for any of various applications involving particle analysis.
C12M 1/34 - Measuring or testing with condition measuring or sensing means, e.g. colony counters
C12M 3/00 - Tissue, human, animal or plant cell, or virus culture apparatus
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
49.
APPARATUS, METHODS AND PROCESSES FOR SORTING PARTICLES AND FOR PROVIDING SEX-SORTED ANIMAL SPERM
A system for sorting a mixture of stained particles, including stained particles having a characteristic A and stained particles having a characteristic B, the system comprising: a. a fluid delivery system for delivering a fluid containing the stained particles in a flow path; b. an electromagnetic radiation source for exciting fluorescence emissions from the stained particles having characteristic A and the stained particles having characteristic B in the flow path; c. a photodetector for detecting the fluorescence emissions from the stained particles; d. a processor in communication with the photodetector for classifying the stained particles according to their fluorescence emissions as either particles having characteristic A or particles having characteristic B; e. a sorting system for sorting the stained particles according to the classification to provide at least one population containing desired particles; f. an analog to digital converter for sampling a time-varying analog output from the photodetector and providing an output including digital information corresponding to the time-varying analog output wherein the time-varying analog output and the corresponding digital information are indicative of characteristic A or characteristic B; and g. a digital signal processor for analyzing and classifying the digital information and providing a sorting signal to the sorting system as a function of the analyzed and classified digital information.
A process for evaluating a set of conditions for staining a population of cells for sorting, the population comprising a first type and a second type of cell, the process comprising: (a) staining a fraction of the population of cells with a fluorescent dye under a set of staining conditions; (b) exposing the stained cells to electromagnetic radiation as the stained cells are passed through an interrogation location of a flow cytometer at a rate, R; (c) determining a fluorescence emission characteristic of the exposed cells; (d) using the determined fluorescence characteristic to classify the exposed cells into two or more sub-populations of cells, one of the sub-populations being an enriched sub-population of the first cell type; (e) determining a coefficient of variation for the fluorescence emission characteristic of the cells of the enriched sub-population to provide an indication of sorting efficiency for the staining conditions; and (f) determining whether to modify any staining condition under which cells are to be stained or the rate, R, at which the stained cells are to be passed through the interrogation location of the flow cytometer in order to improve sorting efficiency.
B07C 5/34 - Sorting according to other particular properties
C12M 1/34 - Measuring or testing with condition measuring or sensing means, e.g. colony counters
C12M 3/00 - Tissue, human, animal or plant cell, or virus culture apparatus
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
G01N 15/00 - Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
A system for sorting a mixture of stained particles having characteristics A and B, having a fluid delivery system for delivering fluid containing stained particles; an electromagnetic radiation source exciting fluorescence emissions from the particles; a photodetector detecting the fluorescence emissions from the particles; a processor classifying the particles according to fluorescence emissions as having characteristic A or B; a sorting system sorting the particles according to classification to provide a population containing desired particles; an analog to digital converter for sampling a time-varying analog output from the photodetector and providing an output including digital information corresponding to the time-varying analog output wherein the time-varying analog output and the corresponding digital information are indicative of characteristic A or B; and a digital signal processor for analyzing and classifying the digital information and providing a sorting signal to the sorting system.
Process for evaluating a set of conditions for staining a population of cells for sorting, the population comprising first and second cell types, the process comprising: (a) staining cells with a fluorescent dye under a set of staining conditions; (b) exposing the stained cells to electromagnetic radiation as the cells pass through an interrogation location of a flow cytometer at a rate, R; (c) determining a fluorescence emission characteristic of the cells; (d) using the characteristic to classify the cells into sub-populations of cells, including an enriched sub- population of the first cell type; (e) determining a coefficient of variation for the fluorescence emission characteristic of the cells of the enriched sub-population to provide an indication of sorting efficiency for the staining conditions; and (f) determining whether to modify any staining condition or the rate, R, at which the cells are passed through the interrogation location to improve sorting efficiency.
C12M 1/34 - Measuring or testing with condition measuring or sensing means, e.g. colony counters
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
C12Q 1/04 - Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
patents
