Histometrics: Improvement of the dynamic range of fluorescently stained proteins resolved in electrophoretic gels using hyperspectral imaging

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dc.contributor.author Woodward, Andrew M.
dc.contributor.author Kaderbhai, Mustak Ali
dc.contributor.author Rowland, Jeremy John
dc.contributor.author Kaderbhai, Naheed Nazly
dc.contributor.author Shaw, Adrian David
dc.contributor.author Kell, Douglas B.
dc.date.accessioned 2009-11-10T09:13:37Z
dc.date.available 2009-11-10T09:13:37Z
dc.date.issued 2001-11-21
dc.identifier.citation Woodward , A M , Kaderbhai , M A , Rowland , J J , Kaderbhai , N N , Shaw , A D & Kell , D B 2001 , ' Histometrics: Improvement of the dynamic range of fluorescently stained proteins resolved in electrophoretic gels using hyperspectral imaging ' Proteomics , vol 1 , no. 11 , pp. 1351-1358 . , 10.1002/1615-9861(200111)1:11<1351::AID-PROT1351>3.0.CO;2-0 en
dc.identifier.issn 1615-9861
dc.identifier.other PURE: 127404
dc.identifier.other dspace: 2160/3464
dc.identifier.uri http://hdl.handle.net/2160/3464
dc.description Woodward, A. M., Kaderbhai, N., Kaderbhai, M., Shaw, A., Rowland, J., Kell, D. B. (2001). Histometrics: Improvement of the dynamic range of fluorescently stained proteins resolved in electrophoretic gels using hyperspectral imaging.   Proteomics, 1, (11), 1351-1358. Sponsorship: BBSRC en
dc.description.abstract Most image-based analyses, using absorbance or fluorescence of the spatial distribution of identifiable structures in complex biological systems, use only a very small number of dimensions of possible spectral data for the generation and interpretation of the image. We here extend the concepts of hyperspectral imaging, being developed in remote sensing, into analytical biotechnology. The massive volume of information contained in hyperspectral spectroscopic images requires multivariate analysis in order to extract the chemical and spatial information contained within the data. We here describe the use of multivariate statistical methods to map and quantify common protein staining fluorophores (SYPRO Red, Orange and Tangerine) in electrophoretic gels. Specifically, we find (a) that the `background underpinning limits of detection is due more to proteins that have not migrated properly than to impurities or to ineffective destaining, (b) the detailed mechanisms of staining of SYPRO red and orange are apparently not identical, and in particular (c) that these methods can provide two orders of magnitude improvement in the detection limit per pixel, to levels well below the limit observable optically. en
dc.format.extent 8 en
dc.language.iso eng
dc.relation.ispartof Proteomics en
dc.subject SYPRO stains en
dc.subject Fluorescent staining en
dc.subject Gel electrophoresis en
dc.subject Hyperspectral imaging en
dc.subject Protein quantification en
dc.title Histometrics: Improvement of the dynamic range of fluorescently stained proteins resolved in electrophoretic gels using hyperspectral imaging en
dc.type Text en
dc.type.publicationtype Article (Journal) en
dc.identifier.doi http://dx.doi.org/10.1002/1615-9861(200111)1:11<1351::AID-PROT1351>3.0.CO;2-0
dc.contributor.institution Institute of Biological, Environmental and Rural Sciences en
dc.contributor.institution Department of Computer Science en
dc.description.status Peer reviewed en


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