Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals

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Intravital fluorescence microscopy has emerged as a powerful technique to visualize cellular processes in vivo. However, owing to their size, the objective lenses required have limited physical accessibility to various tissue sites in the internal organs of small animals. The use of small-diameter probes using graded-index (GRIN) lenses expands the capabilities of conventional intravital microscopes to minimally invasive imaging of internal organs. In this protocol, we describe the detailed steps for the fabrication of front-and side-view GRIN probes and the integration and operation of the probes in a confocal microscope to enable visualization of fluorescent cells and microvasculature in various mouse organs. Some experience in building an optical setup is required to complete the protocol. We also present longitudinal imaging of immune cells in renal allografts and tumor development in the colon. Fabrication and integration can be completed in 5-7 h, and a typical in vivo imaging session takes 1-2 h.
Publisher
NATURE PUBLISHING GROUP
Issue Date
2012-08
Language
English
Article Type
Article
Keywords

ENDOSCOPY; MICE; ENDOMICROSCOPY; MICROENDOSCOPY; RESOLUTION; CANCER; CELLS

Citation

NATURE PROTOCOLS, v.7, no.8, pp.1456 - 1469

ISSN
1754-2189
DOI
10.1038/nprot.2012.078
URI
http://hdl.handle.net/10203/103797
Appears in Collection
NT-Journal Papers(저널논문)
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