title: Use of ImageJ to recover information from individual cells in a G protein-coupled receptor assay.
abstract: Live-cell assays used in GPCR research often rely on fluorescence techniques that generate large amounts of raw image data. Consequently, the capacity to accurately and timely extract useful information from image and video data has become more and more important. Image J is an open-source program that provides powerful tools with a simple interface designed to fit the needs of image analysis of most researchers. In this chapter, Image J routines to extract information from individual cells in a calcium GPCR assay are described. In these routines, individual cells in the same image/video data can be separated using either a progressive threshold or a local threshold method. Both methods can be optimized to either a maximum number of selection or maximum area selected resulting in conceptually distinct selections.
authors.affiliation: IBB - Institute for Biotechnology and Bioengineering, Lisbon, Portugal.
authors.forename: João R C
authors.initials: JR
authors.lastname: Trabuco
authors.forename: Sofia Aires M
authors.initials: SA
authors.lastname: Martins
authors.forename: Duarte Miguel F
authors.initials: DM
authors.lastname: Prazeres
mesh_terms: D000814:Aniline Compounds; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002217:Carbachol / Q000494:pharmacology; D005453:Fluorescence; D005456:Fluorescent Dyes; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000706:statistics & numerical data*; D043643:Receptor, Muscarinic M1 / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods / Q000706:statistics & numerical data; D012984:Software; D014741:Video Recording; D014966:Xanthenes
chemical_list: D000814:Aniline Compounds; C409648:Fluo 4; D005456:Fluorescent Dyes; D043643:Receptor, Muscarinic M1; D014966:Xanthenes; D002217:Carbachol; D002118:Calcium