Three-dimensional visualization of microvessel architecture of whole-mount tissue by confocal microscopy.

Three-dimensional visualization of microvessel architecture of whole-mount tissue by confocal microscopy. Research Abstract Details 

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Three-dimensional visualization of microvessel architecture of whole-mount tissue by confocal microscopy. Abstract Text:

R Dickie,R M Bachoo,M A Rupnick,S M Dallabrida,G M Deloid,J Lai,R A Depinho,R A Rogers,

The three-dimensional architecture of the nascent microvascular network is a critical determinant of vascular perfusion in the setting of regenerative growth, vasculopathies and cancer. Current methods for microvessel visualization are limited by insufficient penetration and instability of endothelial immunolabels, inadequate vascular perfusion by the high-viscosity polymers used for vascular casting, and destruction of tissue stroma during the processing required for scanning electron microscopy. The aim of this study was to develop whole-mount tissue processing methods for 3D in situ visualization of the microvasculature that were also compatible with supplementary labeling for other structures of interest in the tissue microenvironment. Here, we present techniques that allow imaging of the microvasculature by confocal microscopy, to depths of up to 1500 mum below the specimen surface. Our approach includes labeling luminal surfaces of endothelial cells by i.v. injection of fluorescently conjugated lectin and filling the microvasculature with carbon or fluorescent nanoparticles/Mercox, followed by optical clearing of thick tissue sections to reduce light scatter and permit 3D visualization of microvessel morphology deep into the sample. Notably, tissue stroma is preserved, allowing simultaneous labeling of other structures by immunohistochemistry or nuclear dyes. Results are presented for various murine tissues including fat, muscle, heart and brain under conditions of normal health, as well as in the setting of a glioma model growing in the subcutaneous space or orthotopically in the brain parenchyma.

Three-dimensional visualization of microvessel architecture of whole-mount tissue by confocal microscopy. Publishing Authors By Initials

R Dickie,RM Bachoo,MA Rupnick,SM Dallabrida,GM Deloid,J Lai,RA Depinho,RA Rogers,

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Three-dimensional visualization of microvessel architecture of whole-mount tissue by confocal microscopy. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Microvascular research

VOLUME: 72

Page Numbers: 20-6

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ISSN: 0026-2862

DAY: 23

MONTH: 06

YEAR: 2006

Three-dimensional visualization of microvessel architecture of whole-mount tissue by confocal microscopy. Information

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LANGUAGE: eng

NlmUniqueID: 165035

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AFFILIATION: Biomedical Imaging Laboratory, Department of Environmental Health, Harvard School of Public Health, Boston, MA 02115, USA. rdickie@hsph.harvard.edu

Country: United States

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MEDLINETA: Microvasc Res

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