Automated microinjection for zebrafish xenograft models

npj Biomedical Innovations., vol. 2

Abstract

Zebrafish xenograft models are increasingly recognized for predicting patient responses to cancer therapeutics, suggesting their potential as clinical diagnostic tools. However, precise microinjection of cancer cells into numerous small and fragile zebrafish larvae is laborious, requires extensive training for new operators, and often yields variable results, limiting their clinical and drug discovery applications. To address these challenges, we have designed, built, and validated an automated microinjection robot. The robot performs injections into the vasculature, perivitelline space, and hindbrain ventricle in both fully automated and semi-automated modes. Combined results demonstrate an average injection success rate of approximately 60% and larvae survival exceeding 70%, comparable to manual methods, with the fully automated mode being twice as fast. This automation reduces the need for extensive personnel training while enhancing reproducibility, efficiency, and accuracy, paving the way for more extensive use of zebrafish xenograft models in drug discovery and patient diagnostics.

Authors 20

1. Yi Ding
   Affiliation as printed

   Life Science Methods BV, Leiden, the Netherlands

2. Kees-Jan van der Kolk
   Affiliation as printed

   Life Science Methods BV, Leiden, the Netherlands

3. Wietske van der Ent

   University of Oslo

   Affiliation as printed

   Centre for Molecular Medicine Norway (NCMM), University of Oslo, Oslo, Norway

4. Michele Scotto di Mase

   ZeClinics (Spain)

   Affiliation as printed

   ZeClinics SL, Barcelona, Spain

5. Saskia Kowald

   BioReperia AB (Sweden)

   Affiliation as printed

   BioReperia AB, Linköping, Sweden

6. Jenny Huizing Aachen Oncode Institute Department Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Oncode Institute, Dept. Cell & Chemical Biology, Leiden University Medical Center, Leiden, the Netherlands

7. Jana M. Vidal Teuton Aachen Oncode Institute Department Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Oncode Institute, Dept. Cell & Chemical Biology, Leiden University Medical Center, Leiden, the Netherlands

8. Gunja Mishra Aachen Oncode Institute Department Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Oncode Institute, Dept. Cell & Chemical Biology, Leiden University Medical Center, Leiden, the Netherlands

9. Maxime Kempers
   Affiliation as printed

   Life Science Methods BV, Leiden, the Netherlands

10. Rusul Almter

    BioReperia AB (Sweden)

    Affiliation as printed

    BioReperia AB, Linköping, Sweden

11. Sandra Kunz

    University of Oslo

    Affiliation as printed

    Centre for Molecular Medicine Norway (NCMM), University of Oslo, Oslo, Norway

12. Laurine Munier

    University of Oslo

    Affiliation as printed

    Centre for Molecular Medicine Norway (NCMM), University of Oslo, Oslo, Norway

13. Carl Johannes Koschmann

    University of Michigan

    Affiliation as printed

    Department of Pediatrics, University of Michigan, Ann Arbor, MI, USA

14. Sebastian M. Waszak

    University of Oslo

    Affiliation as printed

    Centre for Molecular Medicine Norway (NCMM), University of Oslo, Oslo, Norway

15. Vincenzo Di Donato

    ZeClinics (Spain)

    Affiliation as printed

    ZeClinics SL, Barcelona, Spain

16. Sylvia Dyballa

    ZeClinics (Spain)

    Affiliation as printed

    ZeClinics SL, Barcelona, Spain

17. Peter ten Dijke Aachen Oncode Institute Department Cell and Chemical Biology

    Leiden University Medical Center

    Affiliation as printed

    Oncode Institute, Dept. Cell & Chemical Biology, Leiden University Medical Center, Leiden, the Netherlands

18. Camila Vicencio Esguerra

    University of Oslo

    Affiliation as printed

    Centre for Molecular Medicine Norway (NCMM), University of Oslo, Oslo, Norway

19. Lasse D. Jensen

    Linköping University · BioReperia AB (Sweden)

    Affiliation as printed

    BioReperia AB, Linköping, Sweden

    Division of Diagnostics and Specialist Medicine, Linköping University, Linköping, Sweden

20. Jan de Sonneville corresponding Aachen Life Science Methods BV

    Leiden University Medical Center

    Affiliation as printed

    Life Science Methods BV, Leiden, the Netherlands. jan@lifesciencemethods.com