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SOFTWARE AND METHODS

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Software

Stefania Marcotti, PhD
https://github.com/stemarcotti/PIV
Particle Image Velocimetry (PIV) package developed in the Stramer Lab (King's College London, UK). Please see our publication on Nature Cell Biology (https://www.nature.com/articles/s41556-019-0411-5) for more details.
If you are interested in using this analysis tool on your data, please do get in touch with Brian or Stefania (see People and Contact pages for details)! We have extensive experience in adjusting the parameters for different kind of input movies to ensure that the analysis returns sensible results, and we will be more than happy to help.

https://github.com/stemarcotti/Fiji-macros
Miscellaneous of Fiji macros which might be helpful for batch analysis of images and movies (think saving all your open images, or run simple cell nuclei segmentation).
Do get in touch should you have any question or should anything not work as expected!

Methods
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Working with the Drosophila embryo

In our lab, many projects use Drosophila embryogenesis to study the role and regulation of cell migration in animal physiology, and mechanisms controlling extracellular matrix formation. To look at the fly embryo in real time, we have developed a variety of live imaging techniques, using a variety of microscopy systems. Here, there are some links to our more frequently used protocols, from embryo collection to imaging.
Drosophila embryo collection
  • protocols.io
Drosophila egg dechorionation
  • protocols.io
Live dissection of Drosophila embryos
  • Peter Lee HK, Wright AP, Zinn K. (2009) Live dissection of Drosophila embryos: streamlined methods for screening mutant collections by antibody staining. J Vis Exp. Dec 29;(34):1647.
Live imaging of the Drosophila embryo
  • Matsubayashi Y, Louani A, Dragu A, Sanchez-Sanchez B, Serna-Morales E, Yolland L, Gyoergy A, Vizcay G, Fleck R, Heddleston J, Chew T, Siekhaus D, Stramer B. (2017) A moving source of matrix components is essential for de novo basement membrane formation. Curr. Biol. 27(22):3526-3534.
  • Sanchez-Sanchez B, Urbano J, Comber K, Dragu A, Wood W, Stramer B, Martin-Bermudo D. (2017) Drosophila embryonic hemocytes use autocrine laminins to strengthen their migratory response.  Cell Rep. 21(6):1461-1470.
  • Evans I, Zanet J, Wood W, Stramer B.  (2010)  Live imaging of Drosophila melanogaster embryonic hemocyte migrations.  J. Vis. Exp  Feb 12;(36) 1696.
  • Stramer B and Wood W. (2009)  Inflammation and wound healing in Drosophila. Methods in Molecular Biology. 571; 137-49.
Fluorescence recovery after photobleaching (FRAP) in the Drosophila embryo
  • Matsubayashi Y, Louani A, Dragu A, Sanchez-Sanchez B, Serna-Morales E, Yolland L, Gyoergy A, Vizcay G, Fleck R, Heddleston J, Chew T, Siekhaus D, Stramer B. (2017) A moving source of matrix components is essential for de novo basement membrane formation. Curr. Biol. 27(22):3526-3534.
Electron Microscopy (EM) in the Drosophila embryo
  • Matsubayashi Y, Louani A, Dragu A, Sanchez-Sanchez B, Serna-Morales E, Yolland L, Gyoergy A, Vizcay G, Fleck R, Heddleston J, Chew T, Siekhaus D, Stramer B. (2017) A moving source of matrix components is essential for de novo basement membrane formation. Curr. Biol. 27(22):3526-3534.

Stramer Lab

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King’s College London
Randall Centre for Cell & Molecular Biophysics
New Hunt's House, Guy's Campus
London SE1 1UL
United Kingdom

Professor Brian Stramer
​Phone:+44 (0) 207 848 6272
e-mail: brian.m.stramer@kcl.ac.uk

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