Colocalization of fluorescent markers in confocal microscope images of plant cells - PubMed (original) (raw)

Colocalization of fluorescent markers in confocal microscope images of plant cells

Andrew P French et al. Nat Protoc. 2008.

Abstract

This protocol describes the steps needed to perform quantitative statistical colocalization on two-color confocal images, specifically of plant cells. The procedure includes a calibration test to check the chromatic alignment of the confocal microscope. A software tool is provided to calculate the Pearson and Spearman correlation coefficients ('Pearson-Spearman correlation colocalization' ImageJ plug-in) across regions of interest within the image. Steps are included to help the user practice using the software. The result is a quantitative estimate of the amount of colocalization in the images. Manual masking takes about 1-15 min per image, depending on the detail required, and calculating the correlation coefficients is almost instantaneous. Examples of suitable dyes for such two-color colocalization include Oregon Green or Alexa Fluor 488 dyes in the green range (excited with 488-nm laser line) and Alexa Fluor 555 dye in the red range (excited with 543-nm laser line).

PubMed Disclaimer

Cited by

Ubiquitin initiates sorting of Golgi and plasma membrane proteins into the vacuolar degradation pathway.
Scheuring D, Künzl F, Viotti C, Yan MS, Jiang L, Schellmann S, Robinson DG, Pimpl P. Scheuring D, et al. BMC Plant Biol. 2012 Sep 12;12:164. doi: 10.1186/1471-2229-12-164. BMC Plant Biol. 2012. PMID: 22970698 Free PMC article.
Involvement of formyl peptide receptors in receptor for advanced glycation end products (RAGE)--and amyloid beta 1-42-induced signal transduction in glial cells.
Slowik A, Merres J, Elfgen A, Jansen S, Mohr F, Wruck CJ, Pufe T, Brandenburg LO. Slowik A, et al. Mol Neurodegener. 2012 Nov 20;7:55. doi: 10.1186/1750-1326-7-55. Mol Neurodegener. 2012. PMID: 23164356 Free PMC article.
GOLGI TRANSPORT 1B Regulates Protein Export from the Endoplasmic Reticulum in Rice Endosperm Cells.
Wang Y, Liu F, Ren Y, Wang Y, Liu X, Long W, Wang D, Zhu J, Zhu X, Jing R, Wu M, Hao Y, Jiang L, Wang C, Wang H, Bao Y, Wan J. Wang Y, et al. Plant Cell. 2016 Nov;28(11):2850-2865. doi: 10.1105/tpc.16.00717. Epub 2016 Nov 1. Plant Cell. 2016. PMID: 27803308 Free PMC article.
The Accessory Protein ORF3 Contributes to Porcine Epidemic Diarrhea Virus Replication by Direct Binding to the Spike Protein.
Kaewborisuth C, He Q, Jongkaewwattana A. Kaewborisuth C, et al. Viruses. 2018 Jul 28;10(8):399. doi: 10.3390/v10080399. Viruses. 2018. PMID: 30060558 Free PMC article.
The host exocyst complex is targeted by a conserved bacterial type-III effector that promotes virulence.
Michalopoulou VA, Mermigka G, Kotsaridis K, Mentzelopoulou A, Celie PHN, Moschou PN, Jones JDG, Sarris PF. Michalopoulou VA, et al. Plant Cell. 2022 Aug 25;34(9):3400-3424. doi: 10.1093/plcell/koac162. Plant Cell. 2022. PMID: 35640532 Free PMC article.

Colocalization of fluorescent markers in confocal microscope images of plant cells - PubMed (original) (raw)