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Bioconjug Chem. 2017 Jan 18;28(1):253-259. doi: 10.1021/acs.bioconjchem.6b00500. Epub 2016 Dec 5.

Exploring Passive Clearing for 3D Optical Imaging of Nanoparticles in Intact Tissues.

Author information

1
Institute of Biomaterials and Biomedical Engineering , Rosebrugh Building, Room 407, 164 College Street, Toronto, Ontario M5S 3G9, Canada.
2
Department of Chemistry, University of Toronto , 80 Saint George Street, Toronto, Ontario M5S 3H6, Canada.
3
Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto , 160 College Street, Room 230, Toronto, Ontario M5S 3E1, Canada.
4
Department of Chemical Engineering, University of Toronto , 200 College Street, Toronto, Ontario M5S 3E5, Canada.
5
Department of Material Science and Engineering, University of Toronto , 160 College Street, Room 450, Toronto, Ontario M5S 3E1, Canada.

Abstract

The three-dimensional (3D) optical imaging of nanoparticle distribution within cells and tissues can provide insights into barriers to nanoparticle transport in vivo. However, this approach requires the preparation of optically transparent samples using harsh chemical and physical methods, which can lead to a significant loss of nanoparticles and decreased sensitivity of subsequent analyses. Here, we investigate the influence of electrophoresis and clearing time on nanoparticle retention within intact tissues and the impact of these factors on the final 3D image quality. Our findings reveal that longer clearing times lead to a loss of nanoparticles but improved transparency of tissues. We discovered that passive clearing improved nanoparticle retention 2-fold compared to results from electrophoretic clearing. Using the passive clearing approach, we were able to observe a small population of nanoparticles undergoing hepatobiliary clearance, which could not be observed in liver tissues that were prepared by electrophoretic clearing. This strategy enables researchers to visualize the interface between nanomaterials and their surrounding biological environment with high sensitivity, which enables quantitative and unbiased analysis for guiding the next generation of nanomedicine designs.

PMID:
27801589
DOI:
10.1021/acs.bioconjchem.6b00500
[Indexed for MEDLINE]

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