Eshre 2011 Paul Knaggs Embryo Biopsy Using Saturn Active Presentation

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Presentation given by Paul Knaggs, IVF Hammersmith at ESHRE 2011, Stockholm, Sweden on the subject of embryo biopsy using Saturn Active presentation

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Eshre 2011 Paul Knaggs Embryo Biopsy Using Saturn Active Presentation

  1. 1. Embryo Biopsy using Saturn Active™ Dr Paul Knaggs Lab Manager IVF Hammersmith
  2. 2. What is Saturn Active™? <ul><li>Microsurgical laser system </li></ul><ul><ul><li>Quick and easy tool for partial or total zona ablation </li></ul></ul><ul><ul><li>Many advantages over chemical or mechanical methods </li></ul></ul><ul><ul><li>Many applications </li></ul></ul>
  3. 3. The Saturn Active™ System <ul><li>Class 1 laser product </li></ul><ul><li>Unique motorised laser module </li></ul><ul><li>Easily calibrated to maintain accuracy </li></ul><ul><li>Integration with Integra TI™ micromanipulation system and Cronus Software </li></ul>
  4. 4. Applications of Saturn Active™ <ul><li>Laser assisted hatching (LAH) </li></ul><ul><li>Oocyte Biopsy </li></ul><ul><li>Cleavage stage embryo biopsy </li></ul><ul><li>Blastocyst Biopsy </li></ul><ul><li>Blastomere collapse </li></ul><ul><li>Sperm immobilisation </li></ul><ul><li>Nuclear transfer </li></ul>
  5. 5. Applications of Saturn Active™ <ul><li>Oocyte polar body Biopsy </li></ul><ul><ul><ul><ul><ul><li>Routine PGS, Maternal translocations (may need both polar bodies for high accuracy) </li></ul></ul></ul></ul></ul><ul><li>Cleavage stage embryo biopsy </li></ul><ul><ul><ul><ul><ul><li>PGD for translocation and single gene disorders (fresh ET) </li></ul></ul></ul></ul></ul><ul><li>Blastocyst Biopsy </li></ul><ul><ul><ul><ul><ul><li>PGD for translocation and single gene disorders/ PGS (may need to cryopreserve for single gene depending on analytical protocol - ie non-CGH) </li></ul></ul></ul></ul></ul>
  6. 6. How is it done? <ul><li>Integra TI™ + Saturn Active ™ laser </li></ul><ul><li>Humagen microtools </li></ul><ul><li>Cook Medium </li></ul>
  7. 7. Procedure - oocytes <ul><li>ICSI at 40 hrs post-hCG </li></ul><ul><li>Biopsy immediately after ICSI </li></ul><ul><li>PBs washed in PBS and transferred to eppendorf with 130  M Cook stripper tip </li></ul><ul><li>Oocytes cultured individually in 12  l drops in 4 well dishes </li></ul>
  8. 8. Technique
  9. 9. Procedure -cleavage stage <ul><li>Translocations </li></ul><ul><li>Single gene disorders </li></ul><ul><li>ICSI at 40 hrs post-hCG </li></ul><ul><li>Biopsy on day 3 of development </li></ul><ul><li>Utilise Ca/Mg free medium </li></ul><ul><li>Blastomeres washed in PBS and transferred to eppendorf with 130  M Cook stripper tip </li></ul><ul><li>Embryos cultured individually in 12  l drops in 4 well dishes </li></ul>
  10. 10. Technique
  11. 11. Procedure -Blastocyst stage <ul><li>Translocations </li></ul><ul><li>Single gene disorders </li></ul><ul><li>ICSI at 40 hrs post-hCG </li></ul><ul><li>Zona breach on day 3 </li></ul><ul><li>Biopsy On day 5 or day 6 </li></ul><ul><li>Trophectoderm washed in PBS and transferred to eppendorf with 130  M Cook stripper tip </li></ul><ul><li>Embryos cultured individually in 12  l drops in 4 well dishes </li></ul>
  12. 12. Technique
  13. 13. Potential problems <ul><li>Laser alignment </li></ul><ul><ul><ul><ul><ul><li>Holes not being where they should </li></ul></ul></ul></ul></ul><ul><li>Damage to oolema </li></ul><ul><ul><ul><ul><ul><li>Easily damaged - may consider changing PB position of pipette type </li></ul></ul></ul></ul></ul><ul><li>Contamination </li></ul><ul><ul><ul><ul><ul><li>Operator or other cells </li></ul></ul></ul></ul></ul><ul><li>Fragmented PBs </li></ul><ul><ul><ul><ul><ul><li>May need to hoover the PV space </li></ul></ul></ul></ul></ul><ul><li>Lysed cells </li></ul><ul><ul><ul><ul><ul><li>Cells without nucleus </li></ul></ul></ul></ul></ul>

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