Since its inception in the field of in vivo imaging, endomicroscopy through optical fiber bundles, or probe-based Confocal
Laser Endomicroscopy (pCLE), has extensively proven the benefit of in situ and real-time examination of living tissues at
the microscopic scale. By continuously increasing image quality, reducing invasiveness and improving system ergonomics,
Mauna Kea Technologies has turned pCLE not only into an irreplaceable research instrument for small animal imaging, but
also into an accurate clinical decision making tool with applications as diverse as gastrointestinal endoscopy, pulmonology
The current implementation of pCLE relies on a single fluorescence spectral band making different sources of in vivo
information challenging to distinguish. Extending the pCLE approach to multi-color endomicroscopy therefore appears
as a natural plan. Coupling simultaneous multi-laser excitation with minimally invasive, microscopic resolution, thin
and flexible optics, allows the fusion of complementary and valuable biological information, thus paving the way to a
combination of morphological and functional imaging.
This paper will detail the architecture of a new system, Cellvizio Dual Band, capable of video rate in vivo and in situ
multi-spectral fluorescence imaging with a microscopic resolution. In its standard configuration, the system simultaneously
operates at 488 and 660 nm, where it automatically performs the necessary spectral, photometric and geometric
calibrations to provide unambiguously co-registered images in real-time. The main hardware and software features, including
calibration procedures and sub-micron registration algorithms, will be presented as well as a panorama of its
current applications, illustrated with recent results in the field of pre-clinical imaging.