Abstract
Visualization of single neurons within their complex environment is a pivotal step towards uncovering the mechanisms that control neural circuit development and function. This chapter provides detailed technical information on how to use Drosophila variants of the mouse Brainbow-2 system, called Flybow, for stochastic labeling of cells with different fluorescent proteins in one sample. We first describe the genetic strategies and the heat shock regime required for induction of recombination events. This is followed by a detailed protocol as to how to prepare samples for imaging. Finally, we provide specifications to facilitate multichannel image acquisition using confocal microscopy.
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Acknowledgements
We thank J. Goedhart for sharing the mTurquoise cDNA. D.H. developed the original FB transgenes, while N.S. generated the second set of FB transgenes and validated the new hs-mFLP5 insertions. The original Flybow approach was developed in collaboration with B.J. Dickson, S. Rotkopf, C. Alexandre, and D.M. Bell. We are grateful to H. Apitz, B. Richier, F. Rodrigues, and S. Schrettenbrunner for critical reading of this manuscript. This work is supported by the Medical Research Council (U117581332).
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Shimosako, N., Hadjieconomou, D., Salecker, I. (2014). Flybow to Dissect Circuit Assembly in the Drosophila Brain. In: Sprecher, S. (eds) Brain Development. Methods in Molecular Biology, vol 1082. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-655-9_4
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DOI: https://doi.org/10.1007/978-1-62703-655-9_4
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Publisher Name: Humana Press, Totowa, NJ
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