| In Vivo Molecular Imaging
CRi, Inc.
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DyCE Your Mice for NIR-Perfect Anatomical
Co-Registration
Protocol Authors: Dr. Richard Levenson, CRi; David T. Lynch, Ph.D., properties will require spectral tools for separating them, the core
CRi; James R. Mansfi eld, CRi; Elizabeth M. C. Hillman, Ph.D., capability of CRi’s Maestro product line. If multiplexing is desired as
Columbia University, NY well in the favored near-infrared (NIR) region, this enforces the need
for multispectral techniques. These techniques enhance the imaging
Fluorescence-based molecular imaging in small animals is having a sensitivity by eliminating the ubiquitous autofl uorescence signals
major impact on drug development and disease research. However, from the targeted marker probes. In addition, CRi’s multispectral
a signifi cant challenge to imaging targeted fl uorescent markers in analysis tools have shown utility in analyzing the data by treating
vivo remains: unless the labeled regions are located superfi cially; the time courses like spectral information.
localization, quantitation and host organ identifi cation are impeded
by the effects of light scattering and absorption. Orthotopic tumor
Reference
and disease models are increasingly preferred over less biologically Elizabeth M. C. Hillman, and Anna Moore. All-optical anatomical co-
relevant subcutaneous xenografts. In such studies, substantial
registration for molecular imaging of small animals using dynamic contrast.
diffi culties are encountered in longitudinal studies where animals are
Nature Photonics, 1:526–530 (2007).
growing and are positioned differently for each measurement. We
offer DyCETM, dynamic contrast enhancement, the imaging advance
that will address these issues and radically advance the utility of in
vivo molecular imaging by providing anatomical co-registration of
labeled markers by exploiting in vivo pharmacokinetics of dyes in
small animals in a simple and inexpensive way.
By acquiring a time-series of optical images following a bolus
injection of an inert near-infrared dye indo cyanine green (ICG),
we can repeatably and accurately delineate the major internal
organs of mice using optical imaging alone. This is possible
because each organ has a characteristic pharmacodynamic uptake
or distribution pattern that can be “illuminated” by the kinetics
of dye passing through it to make it distinguishable from other
structures. Subsequently, spatiotemporal analysis can exploit these
characteristic patterns to allow the body-surface representation of
Figure 1. Composite DyCE image showing antomical mapping of internal organs
each organ to be visualized. These in vivo anatomical maps can in live mouse on left, Digimouse anatomical atlas reference image on right.
be overlaid onto simultaneously acquired images of a targeted
Sample and images courtesy Elizabeth M. C. Hillman, Columbia Univ., NY.
molecular probe (detected and distinguished from the mapping
dye via multispectral imaging techniques) to signifi cantly aid in
identifi cation of the marker’s anatomical and physical location.
DyCE can be applied singly or in combination with functionalized
marker probes. Multispectral capabilities are critical here, since
the validity of the simple optical triangulation method for signal co-
localization requires that the molecular marker probe and the organ-
mapping dye have similar optical properties. Dyes with similar optical
Protocol Guide ı 2008 View entire protocol online at www.biotechniques.com/protocol ı BioTechniques ı 39
CRI Protocol.indd 39 10/26/07 11:29:18 AM
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