3D image analysis of fluorescent drug binding

Miquel, M. R., Segura, V., Ali, Z., D'Ocon, M. P., McGrath, J. C. and Daly, C. J. (2005) 3D image analysis of fluorescent drug binding. Molecular imaging, 40(1), pp. 40-52. (doi: 10.1162/15353500200504172) (PMID:15967125)

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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.

Item Type:Articles
Glasgow Author(s) Enlighten ID:Daly, Professor Craig and McGrath, Professor John
Authors: Miquel, M. R., Segura, V., Ali, Z., D'Ocon, M. P., McGrath, J. C., and Daly, C. J.
College/School:College of Medical Veterinary and Life Sciences > School of Life Sciences
Journal Name:Molecular imaging
Publisher:SAGE Publications
ISSN (Online):1536-0121
Published Online:01 January 2005

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