Development of a new bimodal imaging methodology: a combination of fluorescence microscopy and high-resolution secondary ion mass spectrometry.
In this paper, we present a new experimental methodology to combine mass spectrometry (NanoSIMS) with fluorescence microscopy to provide subcellular information on the location of small molecules in cultured cells. We demonstrate this by comparing the distribution of 5-bromo-2-deoxyuridine in the sa...
| Main Authors: | , , , , , |
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| Format: | Journal article |
| Language: | English |
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2010
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| _version_ | 1797103191950622720 |
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| author | Lau, K Christlieb, M Schröder, M Sheldon, H Harris, A Grovenor, C |
| author_facet | Lau, K Christlieb, M Schröder, M Sheldon, H Harris, A Grovenor, C |
| author_sort | Lau, K |
| collection | OXFORD |
| description | In this paper, we present a new experimental methodology to combine mass spectrometry (NanoSIMS) with fluorescence microscopy to provide subcellular information on the location of small molecules in cultured cells. We demonstrate this by comparing the distribution of 5-bromo-2-deoxyuridine in the same cells given by both NanoSIMS analysis and by fluorescence immunohistochemistry. Fiducial markers in the substrates ensured that the images formed by SIMS mapping of bromine ions could be co-registered exactly with images from fluorescence microscopy. The NanoSIMS was shown to faithfully reproduce the information from fluorescence microscopy, but at a much higher spatial resolution. We then show preliminary SIMS images on the distribution of ATN-224, a therapeutic copper chelator for which there is no fluorescent marker, co-registered with conventional Lysotracker and Hoechst stains on the same cells. |
| first_indexed | 2024-03-07T06:16:36Z |
| format | Journal article |
| id | oxford-uuid:f14941c5-1387-4f24-b721-4942b03b1d28 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T06:16:36Z |
| publishDate | 2010 |
| record_format | dspace |
| spelling | oxford-uuid:f14941c5-1387-4f24-b721-4942b03b1d282022-03-27T11:54:54ZDevelopment of a new bimodal imaging methodology: a combination of fluorescence microscopy and high-resolution secondary ion mass spectrometry.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:f14941c5-1387-4f24-b721-4942b03b1d28EnglishSymplectic Elements at Oxford2010Lau, KChristlieb, MSchröder, MSheldon, HHarris, AGrovenor, CIn this paper, we present a new experimental methodology to combine mass spectrometry (NanoSIMS) with fluorescence microscopy to provide subcellular information on the location of small molecules in cultured cells. We demonstrate this by comparing the distribution of 5-bromo-2-deoxyuridine in the same cells given by both NanoSIMS analysis and by fluorescence immunohistochemistry. Fiducial markers in the substrates ensured that the images formed by SIMS mapping of bromine ions could be co-registered exactly with images from fluorescence microscopy. The NanoSIMS was shown to faithfully reproduce the information from fluorescence microscopy, but at a much higher spatial resolution. We then show preliminary SIMS images on the distribution of ATN-224, a therapeutic copper chelator for which there is no fluorescent marker, co-registered with conventional Lysotracker and Hoechst stains on the same cells. |
| spellingShingle | Lau, K Christlieb, M Schröder, M Sheldon, H Harris, A Grovenor, C Development of a new bimodal imaging methodology: a combination of fluorescence microscopy and high-resolution secondary ion mass spectrometry. |
| title | Development of a new bimodal imaging methodology: a combination of fluorescence microscopy and high-resolution secondary ion mass spectrometry. |
| title_full | Development of a new bimodal imaging methodology: a combination of fluorescence microscopy and high-resolution secondary ion mass spectrometry. |
| title_fullStr | Development of a new bimodal imaging methodology: a combination of fluorescence microscopy and high-resolution secondary ion mass spectrometry. |
| title_full_unstemmed | Development of a new bimodal imaging methodology: a combination of fluorescence microscopy and high-resolution secondary ion mass spectrometry. |
| title_short | Development of a new bimodal imaging methodology: a combination of fluorescence microscopy and high-resolution secondary ion mass spectrometry. |
| title_sort | development of a new bimodal imaging methodology a combination of fluorescence microscopy and high resolution secondary ion mass spectrometry |
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