Imaging through the pupal case of Drosophila melanogaster

Mark B. Keroles, Sonya K. Dusseault, Chuchu Liu, Masood R. Mohammed, Christy M. Vadakkan, Jessica H. Amiel, Samantha N. Abel, Elena R. Bensoussan, Benjamin L. Russell, James Baker

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The longstanding use of Drosophila as a model for cell and developmental biology has yielded an array of tools. Together, these techniques have enabled analysis of cell and developmental biology from a variety of methodological angles. Live imaging is an emerging method for observing dynamic cell processes, such as cell division or cell motility. Having isolated mutations in uncharacterized putative cell cycle proteins it became essential to observe mitosis in situ using live imaging. Most live imaging studies in Drosophila have focused on the embryonic stages that are accessible to manipulation and observation because of their small size and optical clarity. However, in these stages the cell cycle is unusual in that it lacks one or both of the gap phases. By contrast, cells of the pupal wing of Drosophila have a typical cell cycle and undergo a period of rapid mitosis spanning about 20 hr of pupal development. It is easy to identify and isolate pupae of the appropriate stage to catch mitosis in situ. Mounting intact pupae provided the best combination of tractability and durability during imaging, allowing experiments to run for several hours with minimal impact on cell and animal viability. The method allows observation of features as small as, or smaller than, fly chromosomes. Adjustment of microscope settings and the details of mounting, allowed extension of the preparation to visualize membrane dynamics of adjacent cells and fluorescently labeled proteins such as tubulin. This method works for all tested fluorescent proteins and can capture submicron scale features over a variety of time scales. While limited to the outer 20 μm of the pupa with a conventional confocal microscope, this approach to observing protein and cellular dynamics in pupal tissues in vivo may be generally useful in the study of cell and developmental biology in these tissues.

Original languageEnglish (US)
Article numbere51239
JournalJournal of Visualized Experiments
Issue number83
DOIs
StatePublished - Jan 23 2014

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Developmental Biology
Pupa
Cytology
Drosophila melanogaster
Mitosis
Drosophila
Cell Biology
Imaging techniques
Cells
Cell Cycle
Proteins
Mountings
Observation
Cell Cycle Proteins
Microscopes
Tubulin
Tissue
Diptera
Cell Division
Cell Movement

Keywords

  • Basic Protocol
  • Confocal microscopy
  • Drosophila
  • Epithelium
  • In vivo
  • Issue 83
  • Live imaging
  • Metamorphosis
  • Mitosis
  • Wing

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Medicine(all)

Cite this

Keroles, M. B., Dusseault, S. K., Liu, C., Mohammed, M. R., Vadakkan, C. M., Amiel, J. H., ... Baker, J. (2014). Imaging through the pupal case of Drosophila melanogaster. Journal of Visualized Experiments, (83), [e51239]. https://doi.org/10.3791/51239

Imaging through the pupal case of Drosophila melanogaster. / Keroles, Mark B.; Dusseault, Sonya K.; Liu, Chuchu; Mohammed, Masood R.; Vadakkan, Christy M.; Amiel, Jessica H.; Abel, Samantha N.; Bensoussan, Elena R.; Russell, Benjamin L.; Baker, James.

In: Journal of Visualized Experiments, No. 83, e51239, 23.01.2014.

Research output: Contribution to journalArticle

Keroles, MB, Dusseault, SK, Liu, C, Mohammed, MR, Vadakkan, CM, Amiel, JH, Abel, SN, Bensoussan, ER, Russell, BL & Baker, J 2014, 'Imaging through the pupal case of Drosophila melanogaster', Journal of Visualized Experiments, no. 83, e51239. https://doi.org/10.3791/51239
Keroles MB, Dusseault SK, Liu C, Mohammed MR, Vadakkan CM, Amiel JH et al. Imaging through the pupal case of Drosophila melanogaster. Journal of Visualized Experiments. 2014 Jan 23;(83). e51239. https://doi.org/10.3791/51239
Keroles, Mark B. ; Dusseault, Sonya K. ; Liu, Chuchu ; Mohammed, Masood R. ; Vadakkan, Christy M. ; Amiel, Jessica H. ; Abel, Samantha N. ; Bensoussan, Elena R. ; Russell, Benjamin L. ; Baker, James. / Imaging through the pupal case of Drosophila melanogaster. In: Journal of Visualized Experiments. 2014 ; No. 83.
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