title: Simple perfusion apparatus for manipulation, tracking, and study of oocytes and embryos.
abstract: OBJECTIVE
  To develop and implement a device and protocol for oocyte analysis at a single cell level. The device must be capable of high resolution imaging, temperature control, perfusion of media, drugs, sperm, and immunolabeling reagents all at defined flow rates. Each oocyte and resultant embryo must remain spatially separated and defined.
  
  
  METHODS
  Experimental laboratory study.
  
  
  METHODS
  University and academic center for reproductive medicine.
  
  
  METHODS
  Women with eggs retrieved for intracytoplasmic sperm injection (ICSI) cycles, adult female FVBN and B6C3F1 mouse strains, sea stars.
  
  
  METHODS
  Real-time, longitudinal imaging of oocytes after fluorescent labeling, insemination, and viability tests.
  
  
  METHODS
  Cell and embryo viability, immunolabeling efficiency, live cell endocytosis quantification, precise metrics of fertilization, and embryonic development.
  
  
  RESULTS
  Single oocytes were longitudinally imaged after significant changes in media, markers, endocytosis quantification, and development, all with supreme control by microfluidics. Cells remained viable, enclosed, and separate for precision measurements, repeatability, and imaging.
  
  
  CONCLUSIONS
  We engineered a simple device to load, visualize, experiment, and effectively record individual oocytes and embryos without loss of cells. Prolonged incubation capabilities provide longitudinal studies without need for transfer and potential loss of cells. This simple perfusion apparatus provides for careful, precise, and flexible handling of precious samples facilitating clinical IVF approaches.
authors.affiliation: Center for Biomedical Engineering, School of Engineering, Brown University, Providence, Rhode Island.
authors.forename: Stephanie L
authors.initials: SL
authors.lastname: Angione
authors.affiliation: Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, Rhode Island.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Oulhen
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Women & Infants Hospital, Providence, Rhode Island; The Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Lynae M
authors.initials: LM
authors.lastname: Brayboy
authors.affiliation: Center for Biomedical Engineering, School of Engineering, Brown University, Providence, Rhode Island.
authors.forename: Anubhav
authors.initials: A
authors.lastname: Tripathi
authors.affiliation: Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, Rhode Island. Electronic address: rhet@brown.edu.
authors.forename: Gary M
authors.initials: GM
authors.lastname: Wessel
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D058948:Cell Tracking / Q000295:instrumentation*; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005260:Female; D006801:Humans; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008846:Micromanipulation / Q000295:instrumentation*; D008853:Microscopy / Q000295:instrumentation*; D009865:Oocytes / Q000166:cytology*
keywords: Oocyte; cell viability assay; endocytosis activity; hydrodynamic trap; microfluidics