Luorescence signal is determined within handdrawn regions of interest selected to tightly enclose the whole slice of every single glomerulus captured within the imaging plane. DFF is calculated because the ratio of (F F) F,exactly where F may be the instantaneous fluorescence signal and F is calculated as the th percentile of the fluorescence signal inside a sliding frame window. For combining the responses of person flies across animals,we normalized the DFF responses from every single person fly for the th percentile on the DFF across all visual stimuli within a single experiment. All responses are the mean in the imply response (across repeated stimulus presentations) of every single of 5 flies. Error bars indicate imply SEM. All significance results presented for Ca imaging had been determined using the MannWhitney test.Wu et al. eLife ;:e. DOI: .eLife. ofResearch articleNeuroscienceAcknowledgementsWe thank the Janelia FlyLight Project Team and Teri Ngo for support with brain dissections,histology and confocal imaging and Janelia Scientific Computing for image processing and information analysis tools. Heather Dionne produced the pJFRC construct. Yoshi Aso shared an early version with the arena assay and Steve Sawtelle,Igor Negrashov and Jinyang Liu made improvements to this apparatus. JiYoung Kim provided the arena utilised for the stochastic activation experiments. We also thank William Rowell for technical help,James Strother for the Ca imaging computer software,Tom Clandinin for the norpA flies,Zhiyong Liu,Rajyashree Sen,Yoshi Aso and Wyatt Korff for guidance,Kei Ito,Alice Robie,Kristin Branson,Ines Ribeiro and Barry Dickson for communicating unpublished benefits,Axel Borst for comments on the manuscript,Andrew Straw and colleagues for agreeing to a shared nomenclature for the LC cell kinds and Crystal Di Pietro for administrative help. Ming Wu was a Helen Hay Whitney Foundation Postdoctoral Fellow.Further informationFundingFunder Howard Hughes Health-related Institute Author Ming Wu Aljoscha Nern W Ryan Williamson Mai M Morimoto Michael B Reiser Gwyneth M Card Gerald M RubinThe funders had no function in study style,data collection and interpretation,or the choice to submit the operate for publication.Author contributions MW,Conceived the study,Generated the splitGAL lines and created the unilateral activation approach,Performed the behavioral experiments and analyses,Contributed to data analysis and writing of your paper; AN,Conceived the study,Performed the buy MK5435 anatomical characterization in the LC cells,Generated the splitGAL lines and developed the unilateral activation approach,Contributed to data evaluation and writing of your paper; WRW,Performed the behavioral experiments and analyses,Contributed to data analysis and writing from the paper; MMM,Performed the calcium imaging experiments,Contributed to information evaluation and writing from the paper; MBR,Provided input around the calcium imaging experiments,Contributed to data analysis and writing on the paper; GMC,Supplied input around the behavioral experiments and analyses,Contributed to data analysis and writing PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/23880851 of the paper; GMR,Conceived the study,Contributed to data evaluation and writing with the paper Author ORCIDs Ming Wu,http:orcid.org Aljoscha Nern,http:orcid.orgX Michael B Reiser,http:orcid.org Gerald M Rubin,http:orcid.orgAdditional filesSupplementary files . Supplementary file . 4 tables with details on LC neuron anatomy,final results of behavioral experiments and experimental genotypes. (A) Summary with the anatomical properties of the LC neuron types.