Automated microinjection for zebrafish xenograft models

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract Zebrafish xenograft models have been increasingly recognized for their ability to predict patient responses to cancer therapeutics, suggesting their potential as diagnostic tools in clinical settings. However, these models require the precise microinjection of cancer cell suspensions in many small and fragile zebrafish larvae. Manual injections are so challenging that, even after months of training, variability in experimental results persists among researchers. This limits the uptake and deployment of zebrafish xenograft models for clinical use and drug discovery. To address this challenge, we have designed, built, and validated an automated microinjection robot. Combined results of injections into the vasculature, perivitelline space, and hindbrain ventricle demonstrated an average injection success rate of approximately 60%, with a larvae survival rate exceeding 70%, comparable to manual injections using a traditional micromanipulator. Notably, the full automated mode was twice as fast as manual injections. This automation of the microinjection process significantly reduces the need for extensive personnel training while it enhances 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
12. Laurine Munier

    University of Oslo

    Affiliation as printed

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

13. Carl Johannes Koschmann
14. Sebastian M. Waszak
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
    Affiliation as printed

    Life Science Methods BV, Leiden, the Netherlands