Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm

CAFC
  • US 7,723,116 B2
  • Filed: 05/25/2006
  • Issued: 05/25/2010
  • Est. Priority Date: 05/15/2003
  • Status: Active Grant
First Claim
Patent Images

1. A method of flow cytometry sperm processing comprising the steps of:

  • establishing a sheath fluid;

    flowing said sheath fluid into at least one nozzle;

    providing a percentage of a fluorescent dye stain as compared to a standard control, said percentage of stain selected from a group consisting of about 90%, about 80%, about 70%, and about 60% of maximum stain;

    staining sperm cells with said percentage of said fluorescent dye stain to provide irradiatable stained sperm cells;

    injecting said irradiatable stained sperm cells into said sheath fluid;

    utilizing one radiation source;

    splitting a radiation into at least two beams;

    directing each of said beams to said at least one nozzle and said sperm cells;

    multiply subjecting said irradiatable sperm cells to said radiation for a first amount of time;

    multiply terminating said radiation of said irradiatable sperm cells for a second amount of time;

    multiply exciting said irradiatable sperm cells with said radiation;

    wherein said step of multiply subjecting said irradiatable sperm cells to said radiation for said first amount of time and said step of multiply terminating said radiation of said irradiatable sperm cells for said second amount of time and said step of multiply exciting said irradiatable sperm cells with said radiation are performed in a sequential manner then repeated in order at a repetition rate;

    emitting fluorescence from said excited sperm cells;

    detecting an amount of said emitted fluorescence from each of said sperm cells;

    evaluating said amount of emitted fluorescence from each of said sperm cells;

    selecting an electrical condition based on the amount of emitted and detected fluorescence from each of said sperm cells to be associated with each of said sperm cells in said sheath fluid flow;

    charging a stream of said irradiatable sperms cell and sheath fluid based on the amount of emitted and detected fluorescence from each of said sperm cells in said sheath fluid flow;

    forming a charged drop having one of said sperm cells located therein;

    isolating said charged drop from said sheath fluid flow;

    deflecting said charged drops;

    sorting said sperm cells; and

    collecting said sorted sperm cells.

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