Post Biopsy Procedures - Dr Paul Knaggs, Consultant Embryologist, Wales Fertility Institute

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Dr Paul Knaggs, Consultant Embryologist at Wales Fertility Institute presented on Post Biopsy Procedures at the RI Istanbul Conference and Hands on Workshop on the 23rd and 24th January 2014.
He covered the following throughout his presentation:
- Manipulation of embryos and biopsy material
- Transportation of samples to external testing lab
- Preparation of samples if analysed in-house
- Manipulation of Biopsy Samples
- Biopsy documentation and results

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Post Biopsy Procedures - Dr Paul Knaggs, Consultant Embryologist, Wales Fertility Institute

  1. 1. Post Biopsy Procedures Dr Paul Knaggs Consultant Embryologist Wales Fertility Institute
  2. 2. What happens after Biopsy? • Manipulation of embryos and biopsy material • Labelling of dishes/ drops and sample tubes • Washing of biopsies and placement in tubes • Recording of procedure • Transportation of samples to external testing lab • Courier • Timing of results/ transfer/ cryopreservation • Preparation of samples if analysed in-house • • • • • PCR – real time, QF-PCR Sequencing FISH Array cGH SNP array • Interpretation of results
  3. 3. Manipulation of Biopsy Samples • Limit number of biopsy and wash drops in each biopsy dish • ?1 dish per biopsy • 1 embryologist to biopsy, 1 to tube • Wear gloves, mask, cover arms. • Clean all surfaces and pipettes/ stripper handles Acid Tyrodes (not required for laser) HEPES Biopsy Medium HEPES Post-Biopsy Wash Drops
  4. 4. Manipulation of Biopsy Samples Transfer biopsy with ~100-125 m pipette Embryo wash step (~300 m pipette) Wash dish – PBS Wash dish on unheated stage – no oil overlay Use new pipette for each biopsy transferred to eppendorf (troph biopsy can be sticky) Same pipette can be used for moving embryos
  5. 5. Biopsy Documentation
  6. 6. Biopsy Documentation • Ensure proper completion of documentation • Testing request for genetics lab • Additional lab paperwork for individual embryo culture • Second witness to check transfers of embryos and biopsies to labelled dishes/ tubes • Ensure that the testing lab knows which tests you want and when the samples will be sent • Remember to book a courier if needed • Establish when results will be available • Only way to establish timing of ET or cryo
  7. 7. Preparation of samples for analysis • Many labs now use external testing labs • Samples placed in cold PBS and shipped in labelled PCR-clean tubes • Especially when using PCR and aCGH • Most commercial labs provide kits of reagents/ tubes to ensure quality • If using FISH you may need to fix to slide • Tarkowsky’s fixation method (see Velilla et al. 2002,Reprod Biomed Online 4:210-217). • Need to carefully label the slides with positions of cells
  8. 8. Results • Turn around time will determine if/ embryo transfer is performed • Blastocyst biopsy doesn’t give the testing lab very much time to allow fresh ET
  9. 9. Results PGD outcomes • Normal PGS outcomes • Normal • Affected • Abnormal • Carrier • No Result • ?ET • No Result • ? ET/ ? Re-biopsy

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