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Upright imaging of Drosophila embryos.

Publication ,  Journal Article
Belu, M; Javier, M; Ayasoufi, K; Frischmann, S; Jin, C; Wang, K-C; Sousa-Neves, R; Mizutani, CM
Published in: J Vis Exp
September 13, 2010

Several well-known morphogenetic gradients and cellular movements occur along the dorsal/ventral axis of the Drosophila embryo. However, the current techniques used to view such processes are somewhat limited. The following protocol describes a new technique for mounting fixed and labeled Drosophila embryos for coronal viewing with confocal imaging. This method consists of embedding embryos between two layers of glycerin jelly mounting media, and imaging jelly strips positioned upright. The first step for sandwiching the embryos is to make a thin bedding of glycerin jelly on a slide. Next, embryos are carefully aligned on this surface and covered with a second layer of jelly. After the second layer is solidified, strips of jelly are cut and flipped upright for imaging. Alternatives are described for visualizing the embryos depending upon the type of microscope stand to be used. Since all cells along the dorsal-ventral axis are imaged within a single confocal Z-plane, our method allows precise measurement and comparison of fluorescent signals without photobleaching or light scattering common to 3D reconstructions of longitudinally mounted embryos.

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Published In

J Vis Exp

DOI

EISSN

1940-087X

Publication Date

September 13, 2010

Issue

43

Location

United States

Related Subject Headings

  • Microscopy, Confocal
  • Imaging, Three-Dimensional
  • Glycerol
  • Embryo Culture Techniques
  • Drosophila
  • Animals
  • 3101 Biochemistry and cell biology
  • 1702 Cognitive Sciences
  • 1701 Psychology
  • 0601 Biochemistry and Cell Biology
 

Citation

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Belu, M., Javier, M., Ayasoufi, K., Frischmann, S., Jin, C., Wang, K.-C., … Mizutani, C. M. (2010). Upright imaging of Drosophila embryos. J Vis Exp, (43). https://doi.org/10.3791/2175
Belu, Mirela, Michelle Javier, Katayoun Ayasoufi, Sarah Frischmann, Catherine Jin, Kuo-Chen Wang, Rui Sousa-Neves, and Claudia Mieko Mizutani. “Upright imaging of Drosophila embryos.J Vis Exp, no. 43 (September 13, 2010). https://doi.org/10.3791/2175.
Belu M, Javier M, Ayasoufi K, Frischmann S, Jin C, Wang K-C, et al. Upright imaging of Drosophila embryos. J Vis Exp. 2010 Sep 13;(43).
Belu, Mirela, et al. “Upright imaging of Drosophila embryos.J Vis Exp, no. 43, Sept. 2010. Pubmed, doi:10.3791/2175.
Belu M, Javier M, Ayasoufi K, Frischmann S, Jin C, Wang K-C, Sousa-Neves R, Mizutani CM. Upright imaging of Drosophila embryos. J Vis Exp. 2010 Sep 13;(43).

Published In

J Vis Exp

DOI

EISSN

1940-087X

Publication Date

September 13, 2010

Issue

43

Location

United States

Related Subject Headings

  • Microscopy, Confocal
  • Imaging, Three-Dimensional
  • Glycerol
  • Embryo Culture Techniques
  • Drosophila
  • Animals
  • 3101 Biochemistry and cell biology
  • 1702 Cognitive Sciences
  • 1701 Psychology
  • 0601 Biochemistry and Cell Biology