A novel method for quantified, superresolved, three-dimensional colocalisation of isotropic, fluorescent particles.
Colocalisation, the overlap of subcellular structures labelled with different colours, is a key step to characterise cellular phenotypes. We have developed a novel bioimage informatics approach for quantifying colocalisation of round, blob-like structures in two-colour, highly resolved, three-dimens...
| Publicado en: | Histochemistry & Cell Biology Vol. 139; no. 3; pp. 391 - 403 |
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| Autores principales: | , , , |
| Formato: | Journal Article |
| Publicado: |
Springer Nature
Mar2013
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=104314656&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 104314656 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 09486143 NQI jtl: Histochemistry & Cell Biology issn: 09486143 maglogo: N pubinfo: dt: Mar2013 vid: 139 iid: 3 pid: 237 pub: Springer Nature place: New York, New York artinfo: ui: 104314656 2011936121 10.1007/s00418-012-1068-3 104314656 ppf: 391 ppct: 12 formats: fmt: @attributes: type: P tig: atl: A novel method for quantified, superresolved, three-dimensional colocalisation of isotropic, fluorescent particles. aug: au: Obara, Boguslaw Jabeen, Asma Fernandez, Nelson Laissue, Pierre affil: School of Biological Sciences, University of Essex, Colchester CO4 3SQ UK sug: ab: Colocalisation, the overlap of subcellular structures labelled with different colours, is a key step to characterise cellular phenotypes. We have developed a novel bioimage informatics approach for quantifying colocalisation of round, blob-like structures in two-colour, highly resolved, three-dimensional fluorescence microscopy datasets. First, the algorithm identifies isotropic fluorescent particles, of relative brightness compared to their immediate neighbourhood, in three dimensions and for each colour. The centroids of these spots are then determined, and each object in one location of a colour image is checked for a corresponding object in the other colour image. Three-dimensional distance maps between the centroids of differently coloured spots then display where and how closely they colocalise, while histograms allow to analyse all colocalisation distances. We use the method to reveal sparse colocalisation of different human leukocyte antigen receptors in choriocarcinoma cells. It can also be applied to other isotropic subcellular structures such as vesicles, aggresomes and chloroplasts. The simple, robust and fast approach yields superresolved, object-based colocalisation maps and provides a first indication of protein-protein interactions of fluorescent, isotropic particles. pubtype: Academic Journal doctype: Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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