Fabrication on the microscale: a two-photon polymerized device for oocyte microinjection.
J Assist Reprod Genet
; 39(7): 1503-1513, 2022 Jul.
Article
de En
| MEDLINE
| ID: mdl-35552947
ABSTRACT
PURPOSE:
Intracytoplasmic sperm injection (ICSI) addresses male sub-fertility by injecting a spermatozoon into the oocyte. This challenging procedure requires the use of dual micromanipulators, with success influenced by inter-operator expertise. We hypothesized that minimizing oocyte handling during ICSI will simplify the procedure. To address this, we designed and fabricated a micrometer scale device that houses the oocyte and requires only one micromanipulator for microinjection.METHODS:
The device consisted of 2 components, each of sub-cubic millimeter volume a Pod and a Garage. These were fabricated using 2-photon polymerization. Toxicity was evaluated by culturing single-mouse presumptive zygotes (PZs) to the blastocyst stage within a Pod, with several Pods (and embryos) docked in a Garage. The development was compared to standard culture. The level of DNA damage/repair in resultant blastocysts was quantified (γH2A.X immunohistochemistry). To demonstrate the capability to carry out ICSI within the device, PZs were microinjected with 4-µm fluorescent microspheres and cultured to the blastocyst stage. Finally, the device was assessed for oocyte traceability and high-throughput microinjection capabilities and compared to standard microinjection practice using key parameters (pipette setup, holding then injecting oocytes).RESULTS:
Compared to standard culture, embryo culture within Pods and a Garage showed no differences in development to the blastocyst stage or levels of DNA damage in resultant blastocysts. Furthermore, microinjection within our device removes the need for a holding pipette, improves traceability, and facilitates high-throughput microinjection.CONCLUSION:
This novel device could improve embryo production following ICSI by simplifying the procedure and thus decreasing inter-operator variability.Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Ovocytes
/
Sperme
Limites:
Animals
Langue:
En
Journal:
J Assist Reprod Genet
Sujet du journal:
GENETICA
/
MEDICINA REPRODUTIVA
Année:
2022
Type de document:
Article
Pays d'affiliation:
Australie