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Live Confocal Analysis of Mutant- and Drug-Treated Drosophila Embryos

  • Barbara Fasulo
  • William Sullivan
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1075)

Abstract

The model organism Drosophila melanogaster is particularly well suited for live image analysis. The availability of GFP transgenic flies and a wide array of fluorescent probes, in conjunction with laser scanning confocal microscopy, allow us to image multiple aspects of the cell cycle simultaneously. Confocal microscopy provides the sensitivity and resolution to observe the dynamics of specific cellular events in real time. For example, GFP-histone and rhodamine-labeled tubulin enable one to follow specific nuclear and cytoskeletal events including nuclear envelope formation, nuclear envelope breakdown, spindle formation, centrosome duplication, separation and migration, chromosomes condensation, and segregation. This analysis permits a detailed morphological and temporal description of nuclear and cytoskeletal events in normal or drug-injected embryos.

Key words

Drosophila Microinjection Drugs 

References

  1. 1.
    Minden JS et al (1989) Direct cell lineage analysis in Drosophila melanogaster by time-lapse, three-dimensional optical microscopy of living embryos. J Cell Biol 109(2):505–516PubMedCrossRefGoogle Scholar
  2. 2.
    Kellogg DR, Mitchison TJ, Alberts BM (1988) Behaviour of microtubules and actin filaments in living Drosophila embryos. Development 103(4):675–686PubMedGoogle Scholar
  3. 3.
    Debec A et al (1996) Live analysis of free centrosomes in normal and aphidicolin-treated Drosophila embryos. J Cell Biol 134(1):103–115PubMedCrossRefGoogle Scholar
  4. 4.
    Foe VE, Alberts BM (1983) Studies of nuclear and cytoplasmic behaviour during the five mitotic cycles that precede gastrulation in Drosophila embryogenesis. J Cell Sci 61:31–70PubMedGoogle Scholar
  5. 5.
    Song YH (2005) Drosophila melanogaster: a model for the study of DNA damage checkpoint response. Mol Cells 19(2):167–179PubMedGoogle Scholar
  6. 6.
    Yu KR, Saint RB, Sullivan W (2000) The Grapes checkpoint coordinates nuclear envelope breakdown and chromosome condensation. Nat Cell Biol 2(9):609–615PubMedCrossRefGoogle Scholar
  7. 7.
    Sullivan W et al (1993) Delays in anaphase initiation occur in individual nuclei of the syncytial Drosophila embryo. Mol Biol Cell 4(9):885–896PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Yu KR, Duronio RJ, Sullivan W (1998) Cell cycle checkpoints: safe passage through mitosis. In: Mechanisms of cell division: frontiers in biology. Oxford University Press, OxfordGoogle Scholar
  9. 9.
    Saint R, Patterson B (1993) Zygotic transcription and cell proliferation during Drosophila embryogenesis. Genetica 90(2–3):157–163PubMedCrossRefGoogle Scholar
  10. 10.
    Valdes-Perez RE, Minden JS (1995) Drosophila melanogaster syncytial nuclear divisions are patterned: time-lapse images, hypothesis and computational evidence. J Theor Biol 175(4):525–532PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Barbara Fasulo
    • 1
  • William Sullivan
    • 1
  1. 1.Molecular and Cellular BiologyUniversity of CaliforniaSanta CruzUSA

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