title: Direct embryo tagging and identification system by attachment of biofunctionalized polysilicon barcodes to the zona pellucida of mouse embryos.
abstract: OBJECTIVE
  Is the attachment of biofunctionalized polysilicon barcodes to the outer surface of the zona pellucida an effective approach for the direct tagging and identification of cultured embryos?
  
  
  CONCLUSIONS
  The results achieved provide a proof of concept for a direct embryo tagging system using biofunctionalized polysilicon barcodes, which could help to minimize the risk of mismatching errors (mix-ups) in human assisted reproduction technologies.
  
  
  BACKGROUND
  Even though the occurrence of mix-ups is rare, several cases have been reported in fertility clinics around the world. Measures to prevent the risk of mix-ups in human assisted reproduction technologies are therefore required.
  
  
  METHODS
  Mouse embryos were tagged with 10 barcodes and the effectiveness of the tagging system was tested during fresh in vitro culture (n=140) and after embryo cryopreservation (n = 84). Finally, the full-term development of tagged embryos was evaluated (n =105).
  
  
  METHODS
  Mouse pronuclear embryos were individually rolled over wheat germ agglutinin-biofunctionalized polysilicon barcodes to distribute them uniformly around the ZONA PELLUCIDA surface. Embryo viability and retention of barcodes were determined during 96 h of culture. The identification of tagged embryos was performed every 24 h in an inverted microscope and without embryo manipulation to simulate an automatic reading procedure. Full-term development of the tagged embryos was assessed after their transfer to pseudo-pregnant females. To test the validity of the embryo tagging system after a cryopreservation process, tagged embryos were frozen at the 2-cell stage using a slow freezing protocol, and followed in culture for 72 h after thawing.
  
  
  RESULTS
  Neither the in vitro or in vivo development of tagged embryos was adversely affected. The tagging system also proved effective during an embryo cryopreservation process. Global identification rates higher than 96 and 92% in fresh and frozen-thawed tagged embryos, respectively, were obtained when simulating an automatic barcode reading system, although these rates could be increased to 100% by simply rotating the embryos during the reading process.
  
  
  CONCLUSIONS
  The direct embryo tagging developed here has exclusively been tested in mouse embryos. Its effectiveness in other species, such as the human, is currently being tested.
  
  
  CONCLUSIONS
  The direct embryo tagging system developed here, once tested in human embryos, could provide fertility clinics with a novel tool to reduce the risk of mix-ups in human assisted reproduction technologies.
authors.affiliation: Departament de Biologia Cellular, Fisiologia i Immunologia, Facultat de Biociències, Universitat Autónoma de Barcelona, Bellaterra, Barcelona, Spain.
authors.forename: Sergi
authors.initials: S
authors.lastname: Novo
authors.forename: Oriol
authors.initials: O
authors.lastname: Penon
authors.forename: Leonardo
authors.initials: L
authors.lastname: Barrios
authors.forename: Carme
authors.initials: C
authors.lastname: Nogués
authors.forename: Josep
authors.initials: J
authors.lastname: Santaló
authors.forename: Sara
authors.initials: S
authors.lastname: Durán
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Gómez-Matínez
authors.forename: Josep
authors.initials: J
authors.lastname: Samitier
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Plaza
authors.forename: Luisa
authors.initials: L
authors.lastname: Pérez-García
authors.forename: Elena
authors.initials: E
authors.lastname: Ibáñez
mesh_terms: D000823:Animal Identification Systems; D000818:Animals; D015925:Cryopreservation; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000648:ultrastructure*; D047108:Embryonic Development; D005260:Female; D051379:Mice; D027724:Reproductive Techniques, Assisted; D017655:Silicon Compounds; D015044:Zona Pellucida / Q000648:ultrastructure*
chemical_list: D017655:Silicon Compounds
keywords: assisted reproductive technologies (ART); biofunctionalization; embryo tagging; mix-ups; traceability