Fabrication on the microscale: a two-photon polymerized device for oocyte microinjection

Suliman H. Yagoub; Jeremy G. Thompson; Antony Orth; Kishan Dholakia; Brant C. Gibson; Kylie R. Dunning

doi:10.1007/s10815-022-02485-1

Fabrication on the microscale: a two-photon polymerized device for oocyte microinjection

Suliman H. Yagoub, Jeremy G. Thompson, Antony Orth, Kishan Dholakia, Brant C. Gibson, Kylie R. Dunning^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Number of pages	11
Journal	Journal of Assisted Reproduction and Genetics
Volume	First Online
DOIs	https://doi.org/10.1007/s10815-022-02485-1
Publication status	Published - 12 May 2022

Keywords

ICIS
IVF
Infertility
ART
3D fabrication
High-throughput microinjection

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Yagoub_2022_JARG_FabricationOnTheMicroscale_CC
Copyright © The Author(s) 2022. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
Final published version, 1.32 MBLicence: CC BY

Resonant and shaped photonics for under: Resonant and shaped photonics for understanding the physical and biomedical world
Dholakia, K. (PI) & Gather, M. C. (CoI)
1/08/17 → 31/07/22
Project: Standard

Cite this

@article{51899acd58dd4ba2a8fa98859b157d8b,

title = "Fabrication on the microscale: a two-photon polymerized device for oocyte microinjection",

abstract = "PurposeIntracytoplasmic 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.MethodsThe 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).ResultsCompared 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.ConclusionThis novel device could improve embryo production following ICSI by simplifying the procedure and thus decreasing inter-operator variability.",

keywords = "ICIS, IVF, Infertility, ART, 3D fabrication, High-throughput microinjection",

author = "Yagoub, {Suliman H.} and Thompson, {Jeremy G.} and Antony Orth and Kishan Dholakia and Gibson, {Brant C.} and Dunning, {Kylie R.}",

note = "Open Access funding enabled and organized by CAUL and its Member Institutions. KRD is supported by a Mid-Career Fellowship from the Hospital Research Foundation (C-MCF-58–2019). KD acknowledges funding from the UK Engineering and Physical Sciences Research Council (grant EP/P030017/1). This study was funded by the Australian Research Council (ARC) Centre of Excellence for Nanoscale BioPhotonics (CE140100003).",

year = "2022",

month = may,

day = "12",

doi = "10.1007/s10815-022-02485-1",

language = "English",

volume = "First Online",

journal = "Journal of Assisted Reproduction and Genetics",

issn = "1573-7330",

publisher = "Springer",

}

TY - JOUR

T1 - Fabrication on the microscale

T2 - a two-photon polymerized device for oocyte microinjection

AU - Yagoub, Suliman H.

AU - Thompson, Jeremy G.

AU - Orth, Antony

AU - Dholakia, Kishan

AU - Gibson, Brant C.

AU - Dunning, Kylie R.

N1 - Open Access funding enabled and organized by CAUL and its Member Institutions. KRD is supported by a Mid-Career Fellowship from the Hospital Research Foundation (C-MCF-58–2019). KD acknowledges funding from the UK Engineering and Physical Sciences Research Council (grant EP/P030017/1). This study was funded by the Australian Research Council (ARC) Centre of Excellence for Nanoscale BioPhotonics (CE140100003).

PY - 2022/5/12

Y1 - 2022/5/12

N2 - PurposeIntracytoplasmic 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.MethodsThe 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).ResultsCompared 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.ConclusionThis novel device could improve embryo production following ICSI by simplifying the procedure and thus decreasing inter-operator variability.

AB - PurposeIntracytoplasmic 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.MethodsThe 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).ResultsCompared 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.ConclusionThis novel device could improve embryo production following ICSI by simplifying the procedure and thus decreasing inter-operator variability.

KW - ICIS

KW - IVF

KW - Infertility

KW - ART

KW - 3D fabrication

KW - High-throughput microinjection

U2 - 10.1007/s10815-022-02485-1

DO - 10.1007/s10815-022-02485-1

M3 - Article

SN - 1573-7330

VL - First Online

JO - Journal of Assisted Reproduction and Genetics

JF - Journal of Assisted Reproduction and Genetics

ER -

Fabrication on the microscale: a two-photon polymerized device for oocyte microinjection

Abstract

Keywords

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Resonant and shaped photonics for under: Resonant and shaped photonics for understanding the physical and biomedical world

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