3-D Image Analysis of Fluorescent Drug Binding
Fluorescent ligands provide the means of studying receptors in whole tissues using confocal laser scanning microscopy and have advantages over antibody- or non-fluorescence-based method. Confocal microscopy provides large volumes of images to be measured. Histogram analysis of 3-D image volumes is p...
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| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2005-01-01
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| Series: | Molecular Imaging |
| Online Access: | https://doi.org/10.1162/15353500200504172 |
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| _version_ | 1850087376281403392 |
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| author | M. Raquel Miquel Venessa Segura Z. Ali M. P. D'Ocon J. C. McGrath Craig J. Daly |
| author_facet | M. Raquel Miquel Venessa Segura Z. Ali M. P. D'Ocon J. C. McGrath Craig J. Daly |
| author_sort | M. Raquel Miquel |
| collection | DOAJ |
| description | Fluorescent ligands provide the means of studying receptors in whole tissues using confocal laser scanning microscopy and have advantages over antibody- or non-fluorescence-based method. Confocal microscopy provides large volumes of images to be measured. Histogram analysis of 3-D image volumes is proposed as a method of graphically displaying large amounts of volumetric image data to be quickly analyzed and compared. The fluorescent ligand BODIPY FL-prazosin (QAPB) was used in mouse aorta. Histogram analysis reports the amount of ligand-receptor binding under different conditions and the technique is sensitive enough to detect changes in receptor availability after antagonist incubation or genetic manipulations. QAPB binding was concentration dependent, causing concentration-related rightward shifts in the histogram. In the presence of 10 μM phenoxybenzamine (blocking agent), the QAPB (50 nM) histogram overlaps the autofluorescence curve. The histogram obtained for the 1D knockout aorta lay to the left of that of control and 1B knockout aorta, indicating a reduction in 1D receptors. We have shown, for the first time, that it is possible to graphically display binding of a fluorescent drug to a biological tissue. Although our application is specific to adrenergic receptors, the general method could be applied to any volumetric, fluorescence-image-based assay. |
| format | Article |
| id | doaj-art-24c19afc706e4a1bb1599544efd3f228 |
| institution | DOAJ |
| issn | 1536-0121 |
| language | English |
| publishDate | 2005-01-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Molecular Imaging |
| spelling | doaj-art-24c19afc706e4a1bb1599544efd3f2282025-08-20T02:43:13ZengSAGE PublishingMolecular Imaging1536-01212005-01-01410.1162/1535350020050417210.1162_153535002005041723-D Image Analysis of Fluorescent Drug BindingM. Raquel Miquel0Venessa Segura1Z. Ali2M. P. D'Ocon3J. C. McGrath4Craig J. DalyUniversidad de ValenciaUniversidad de ValenciaUniversity of GlasgowUniversidad de ValenciaUniversity of GlasgowFluorescent ligands provide the means of studying receptors in whole tissues using confocal laser scanning microscopy and have advantages over antibody- or non-fluorescence-based method. Confocal microscopy provides large volumes of images to be measured. Histogram analysis of 3-D image volumes is proposed as a method of graphically displaying large amounts of volumetric image data to be quickly analyzed and compared. The fluorescent ligand BODIPY FL-prazosin (QAPB) was used in mouse aorta. Histogram analysis reports the amount of ligand-receptor binding under different conditions and the technique is sensitive enough to detect changes in receptor availability after antagonist incubation or genetic manipulations. QAPB binding was concentration dependent, causing concentration-related rightward shifts in the histogram. In the presence of 10 μM phenoxybenzamine (blocking agent), the QAPB (50 nM) histogram overlaps the autofluorescence curve. The histogram obtained for the 1D knockout aorta lay to the left of that of control and 1B knockout aorta, indicating a reduction in 1D receptors. We have shown, for the first time, that it is possible to graphically display binding of a fluorescent drug to a biological tissue. Although our application is specific to adrenergic receptors, the general method could be applied to any volumetric, fluorescence-image-based assay.https://doi.org/10.1162/15353500200504172 |
| spellingShingle | M. Raquel Miquel Venessa Segura Z. Ali M. P. D'Ocon J. C. McGrath Craig J. Daly 3-D Image Analysis of Fluorescent Drug Binding Molecular Imaging |
| title | 3-D Image Analysis of Fluorescent Drug Binding |
| title_full | 3-D Image Analysis of Fluorescent Drug Binding |
| title_fullStr | 3-D Image Analysis of Fluorescent Drug Binding |
| title_full_unstemmed | 3-D Image Analysis of Fluorescent Drug Binding |
| title_short | 3-D Image Analysis of Fluorescent Drug Binding |
| title_sort | 3 d image analysis of fluorescent drug binding |
| url | https://doi.org/10.1162/15353500200504172 |
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