Oral Virtual Presentation (Virtual only) ESA-SRB-ANZBMS 2021

Fabrication on the microscale: two-photon polymerised device for microinjection (#157)

Suliman H Yagoub 1 2 3 4 , Brant C Gibson 2 5 , Jeremy G Thompson 1 6 , Kylie R Dunning 1 2 3 4
  1. University of Adelaide, Adelaide, SA, Australia
  2. Australian Research Council (ARC) Centre of Excellence for Nanoscale BioPhotonics (CNBP), Adelaide, SA, Australia
  3. School of Biomedicine, Robinson Research Institute (RRI), Adelaide, South Australia, Australia
  4. School of Physcial Sciences, Institute for Photonics and Advanced Sensing (IPAS), Adelaide, SA, Australia
  5. School of Science, RMIT University, Melbourne, VIC, Australia
  6. Fertilis Pty Ltd, Adelaide, SA, Australia

Intracytoplasmic sperm injection (ICSI) addresses male sub-fertility by injecting a spermatozoon into the oocyte to induce fertilisation. It is a technically challenging procedure that requires dual micromanipulation. In the IVF clinic, only highly skilled embryologists perform ICSI. However, variability in success exists between embryologists within and across clinics. Poor success may be the result of the procedure causing mechanical stress on the oocyte, leading to impaired fertilisation and embryo development. We hypothesise that minimising oocyte handling during ICSI will simplify the procedure and thus improve embryo production. To address this, we designed and fabricated a micrometre-scale transparent device to house the oocyte. The device avoids the use of a holding pipette, requiring only one micromanipulator to perform microinjection of sperm. Computer aided design software was used to design the device, which consisted of 2 components, Pod (670 x 235 x 353 µm; l x w x h) and Garage (1150 x 450 x 345 µm). The fabrication of the device was performed by the optical method of 2-photon polymerisation and houses individual oocytes within a single Pod. We show that an array of Pods docked within a Garage enabled high-throughput microinjection of multiple oocytes. Importantly, the Pod/Garage is embryo safe, demonstrated by the gold-standard mouse embryo assay. The use of the Pod/Garage array for ICSI enabled rapid tracing of oocytes that had already undergone, vs. those that still required microinjection. Finally, we demonstrate that subsequent embryo development can be carried out in the same device with development to blastocyst not significantly different to standard microdrop culture (87.4% ± 3.0% vs 85.3% ± 7.1%, respectively, P > 0.05). This work could improve embryo production by removing intra-operator variability and may form a precursor to automated ICSI.