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.
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References
Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez J-Y, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682
Schneider CA, Rasband WS, Eliceiri KW (2012) Historical commentary NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9:671–676
Zhang R, Xie X (2012) Tools for GPCR drug discovery. Acta Pharmacol Sin 33:372–384
Martins SAM, Trabuco JRC, Monteiro GA, Chu V, Conde JP, Prazeres DMF (2012) Towards the miniaturization of GPCR-based live-cell screening assays. Trends Biotechnol 30:566–574
Liu K, Southall N, Titus SA, Inglese J, Eskay RL, Shinn P, Austin CP, Heilig MA, Zheng W (2010) A multiplex calcium assay for identification of GPCR agonists and antagonists. Assay Drug Dev Technol 8:367–379
Eliceiri KW, Berthold MR, Goldberg IG, Ibáñez L, Manjunath BS, Martone ME, Murphy RF, Peng H, Plant AL, Roysam B, Stuurman N, Stuurmann N, Swedlow JR, Tomancak P, Carpenter AE (2012) Biological imaging software tools. Nat Methods 9:697–710
de Chaumont F, Dallongeville S, Chenouard N, Hervé N, Pop S, Provoost T, Meas-Yedid V, Pankajakshan P, Lecomte T, Le Montagner Y, Lagache T, Dufour A, Olivo-Marin J-C (2012) Icy: an open bioimage informatics platform for extended reproducible research. Nat Methods 9:690–696
Kankaanpää P, Paavolainen L, Tiitta S, Karjalainen M, Päivärinne J, Nieminen J, Marjomäki V, Heino J, White DJ (2012) BioImageXD: an open, general-purpose and high-throughput image-processing platform. Nat Methods 9:683–689
Cardona A, Tomancak P (2012) Current challenges in open-source bioimage informatics. Nat Methods 9:661–665
Martins SAM, Moulas G, Trabuco JRC, Monteiro GA, Chu V, Conde JP, Prazeres DMF (2014) Monitoring intracellular calcium in response to GPCR activation using thin-film silicon photodiodes with integrated fluorescence filters. Biosens Bioelectron 52:232–238
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Trabuco, J.R.C., Martins, S.A.M., Prazeres, D.M.F. (2015). Use of ImageJ to Recover Information from Individual Cells in a G Protein-Coupled Receptor Assay. In: Prazeres, D.M.F., Martins, S.A.M. (eds) G Protein-Coupled Receptor Screening Assays. Methods in Molecular Biology, vol 1272. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-2336-6_11
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DOI: https://doi.org/10.1007/978-1-4939-2336-6_11
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