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Journal of Fluorescence

, Volume 22, Issue 4, pp 1055–1063 | Cite as

Fluorescent Function-Spacer-Lipid Construct Labelling Allows for Real-Time in Vivo Imaging of Cell Migration and Behaviour in Zebrafish (Danio Rerio)

  • Chuan-Ching Lan
  • Deborah Blake
  • Stephen Henry
  • Donald R. LoveEmail author
ORIGINAL PAPER

Abstract

Real-time in vivo imaging of cell migration and behavior has advanced our understanding of physiological processes in situ, especially in the field of immunology. We carried out the transplantation of a mixed population of blood cells from adult zebrafish (Danio rerio) to 2 day old embryos. The blood cells were treated ex vivo with Function-Spacer-Lipid constructs (FSL) incorporating either fluorescein or Atto488 fluorophores (FSL-FLRO4-I or -II). Excellent labeling efficiency was demonstrated by epifluorescence microscopy and FACScan analysis. Real-time video imaging of the recipient fish showed that the functionality of these cells was retained and not affected by the labeling. The usefulness of FSL-FLRO4-I as a contrast agent in microangiography was explored. Overall, we found both FSL-FLRO4-I and-II promising labeling dyes for real-time in vivo imaging in zebrafish.

Keywords

Kodecyte Labeling Fluorescence Zebrafish 

Notes

Acknowledgments

Professor Nicolai Bovin’s team from the Shemyakin Institute of Bioorganic Chemistry, Russian Academy of Sciences, is gratefully acknowledged for preparing FSL-FLRO4-II.

References

  1. 1.
    Lieschke GJ, Currie PD (2007) Animal models of human disease: zebrafish swim into view. Nat Rev Genet 8:353–367PubMedCrossRefGoogle Scholar
  2. 2.
    Lawson ND, Weinstein BM (2002) In vivo imaging of embryonic vascular development using transgenic zebrafish. Dev Biol 248:307–318PubMedCrossRefGoogle Scholar
  3. 3.
    Isogai S, Horiguchi M, Weinstein B (2001) The vascular anatomy of the developing zebrafish: an atlas of embryonic and early larval development. Dev Biol 230:278–301PubMedCrossRefGoogle Scholar
  4. 4.
    Rieger S, Kulkarni RP, Darcy D et al (2005) Quantum dots are powerful multipurpose vital labeling agents in zebrafish embryos. Dev Dyn 234:670–681PubMedCrossRefGoogle Scholar
  5. 5.
    Becker HM, Chen M, Hay JB et al (2004) Tracking of leukocyte recruitment into tissues of mice by in situ labeling of blood cells with the fluorescent dye CFDA SE. J Immunol Methods 286:69–78PubMedCrossRefGoogle Scholar
  6. 6.
    Parish CR (1999) Fluorescent dyes for lymphocyte migration and proliferation studies. Immunol Cell Biol 77:499–508PubMedCrossRefGoogle Scholar
  7. 7.
    Blake D, Bovin N, Bess D et al. (2011) FSL Constructs: A simple method for modifying cell/virion surfaces with a range of biological markers without affecting their viability. JoVE (54): 3289Google Scholar
  8. 8.
    Oliver C, Blake D, Henry S (2011) Modeling transfusion reactions and predicting in vivo cell survival with kodecytes. Transfusion 2011(51):1723–1730CrossRefGoogle Scholar
  9. 9.
    Oliver C, Blake D, and Henry S (2011) In vivo neutralization of anti-A and successful transfusion of A antigen incompatible red cells in an animal model. Transfusion In press doi: 10.1111/j.1537-2995.2011.03184.x
  10. 10.
    Hadac E, Federspiel M, Chernyy E et al (2011) Fluorescein and radiolabeled Function-Spacer-Lipid constructs allow for simple in vitro and in vivo bioimaging of enveloped virions. J Virol Methods 176:78–84PubMedCrossRefGoogle Scholar
  11. 11.
    Blake D, Lan C-C, Love D et al (2010) Fluorophore-kodecytes: fluorescent Function-Spacer-Lipid (FSL) modified cells for in vitro and in vivo analyses. FEBS Journal 277:199Google Scholar
  12. 12.
    LeBlanc J, Bowman TV, Zon L (2007) Transplantation of whole kidney marrow in adult zebrafish. JoVE 2:e159Google Scholar
  13. 13.
    Traver D, Paw B, Poss K et al (2003) Transplantation and in vivo imaging of multilineage engraftment in zebrafish bloodless mutants. Nature Immunol 4:1238–1246CrossRefGoogle Scholar
  14. 14.
    Willett C, Cherry J, Steiner L (1997) Characterization and expression of the recombination activating genes (rag1 and rag2) of zebrafish. Immunogenetics 45:394–404PubMedCrossRefGoogle Scholar
  15. 15.
    Lan C-C, Blake D, Henry S et al (2012) Videos relating to Fluorescent Function-Spacer-Lipid construct labeling allows for real-time in vivo imaging of cell migration and behaviour in zebrafish (Danio rerio). http://hdl.handle.net/10292/3475
  16. 16.
    Bertrand J, Kim A, Teng S et al (2008) CD41+ cmyb + precursors colonize the zebrafish pronephros by a novel migration route to initiate adult hematopoiesis. Development 135:853–1862CrossRefGoogle Scholar
  17. 17.
    Langenau D, Ferrando A, Traver D et al (2004) In vivo tracking of T cell development, ablation, and engraftment in transgenic zebrafish. PNAS 101:7369–7374PubMedCrossRefGoogle Scholar
  18. 18.
    Burns C, Zon L (2006) Homing sweet homing: odyssey of hematopoietic stem cells. Immunity 25:859–862PubMedCrossRefGoogle Scholar
  19. 19.
    Mathias J, Dodd M, Walters K (2009) Characterization of zebrafish larval inflammatory macrophages. Dev Comp Immunol 33:1212–1217PubMedCrossRefGoogle Scholar
  20. 20.
    Yoo S, Deng Q, Cavnar P et al (2010) Differential Regulation of Protrusion and Polarity by PI (3) K during Neutrophil Motility in Live Zebrafish. Dev cell 18:226–236PubMedCrossRefGoogle Scholar
  21. 21.
    Hall C, Flores M, Storm T et al (2007) The zebrafish lysozyme C promoter drives myeloid-specific expression in transgenic fish. BMC Dev Biol 7:42PubMedCrossRefGoogle Scholar
  22. 22.
    Mathias J, Perrin B, Liu T et al (2006) Resolution of inflammation by retrograde chemotaxis of neutrophils in transgenic zebrafish. J Leukoc Biol 80:1281–1288PubMedCrossRefGoogle Scholar
  23. 23.
    Niethammer P, Grabher C, Look A et al (2009) A tissue-scale gradient of hydrogen peroxide mediates rapid wound detection in zebrafish. Nature 459:996–999PubMedCrossRefGoogle Scholar
  24. 24.
    White RM, Sessa A, Burke C et al (2008) Transparent adult zebrafish as a tool for in vivo transplantation analysis. Cell Stem Cell 2:83–189CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Chuan-Ching Lan
    • 1
  • Deborah Blake
    • 2
  • Stephen Henry
    • 2
  • Donald R. Love
    • 1
    • 3
    Email author
  1. 1.School of Biological SciencesUniversity of AucklandAucklandNew Zealand
  2. 2.Biotechnology Research InstituteAUT UniversityAucklandNew Zealand
  3. 3.LabPLUSAuckland City HospitalAucklandNew Zealand

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