Luorescence signal is determined inside handdrawn regions of interest chosen to tightly enclose the complete slice of each glomerulus captured inside 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 because the th percentile from the fluorescence signal within a sliding frame window. For combining the responses of person flies across animals,we normalized the DFF responses from every person fly towards the th percentile on the DFF across all visual stimuli within a single experiment. All responses will be the imply of the imply response (across repeated stimulus presentations) of every of 5 flies. Error bars indicate mean SEM. All ALS-8112 significance results presented for Ca imaging were determined using the MannWhitney test.Wu et al. eLife ;:e. DOI: .eLife. ofResearch articleNeuroscienceAcknowledgementsWe thank the Janelia FlyLight Project Group and Teri Ngo for assistance with brain dissections,histology and confocal imaging and Janelia Scientific Computing for image processing and data analysis tools. Heather Dionne made the pJFRC construct. Yoshi Aso shared an early version with the arena assay and Steve Sawtelle,Igor Negrashov and Jinyang Liu produced improvements to this apparatus. JiYoung Kim provided the arena utilized for the stochastic activation experiments. We also thank William Rowell for technical help,James Strother for the Ca imaging software program,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Aso and Wyatt Korff for guidance,Kei Ito,Alice Robie,Kristin Branson,Ines Ribeiro and Barry Dickson for communicating unpublished final results,Axel Borst for comments on the manuscript,Andrew Straw and colleagues for agreeing to a shared nomenclature for the LC cell forms and Crystal Di Pietro for administrative assistance. Ming Wu was a Helen Hay Whitney Foundation Postdoctoral Fellow.Additional informationFundingFunder Howard Hughes Medical Institute Author Ming Wu Aljoscha Nern W Ryan Williamson Mai M Morimoto Michael B Reiser Gwyneth M Card Gerald M RubinThe funders had no role in study style,information collection and interpretation,or the selection to submit the function for publication.Author contributions MW,Conceived the study,Generated the splitGAL lines and created the unilateral activation strategy,Performed the behavioral experiments and analyses,Contributed to data analysis and writing of the paper; AN,Conceived the study,Performed the anatomical characterization of your LC cells,Generated the splitGAL lines and developed the unilateral activation strategy,Contributed to information analysis and writing of your paper; WRW,Performed the behavioral experiments and analyses,Contributed to information analysis and writing of your paper; MMM,Performed the calcium imaging experiments,Contributed to data analysis and writing from the paper; MBR,Offered input on the calcium imaging experiments,Contributed to information evaluation and writing from 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 information evaluation and writing in 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 information on LC neuron anatomy,results of behavioral experiments and experimental genotypes. (A) Summary in the anatomical properties of the LC neuron varieties.