Optical imaging of whole animals or animal organs is a rapidly growing field in translational research revealing the molecular events underlying disease and disease treatment mechanisms in cardiovascular, cancer and neurological research. Here we present a custom-built imaging system for visualisation of 3D distribution of fluorescent markers with high-resolution tissue structure and vasculature network images of small animals or whole organs.
The 3D Fluorescence Imaging Cryomicrotome System (3D-FICS) yields comprehensive structural and functional biological information by combining fluorescence remittance imaging. Moreover, the 3D-FICS is adapted to record large series of high-resolution images (2048 x 2048 pixels, with a selectable resolution of 27, 16 and 8μm, corresponding to a FOV of 53, 31 and 16mm) of the bulk tissue remaining in the cryomicrotome in a fully automated manner. All components are controlled through custom software (Labview) to enable fully automated serial cutting and imaging sessions. To ensure 3D reconstructions with isotropic voxels, the slice thickness of the cryomicrotome is set to match the imaging resolution of the camera.
Wavelength-selective illumination of the tissue is carried out using a Supercontinuum laser in conjunction with a tunable bandpass filter (400nm - 830nm) with a tunable bandwidth (5nm - 100nm) Similarly, a tunable emission filter (440nm - 800nm) with a bandwidth ranging from 11nm to 15nm is used for spectrally-resolved imaging of the fluorescent tracers injected in the specimen.
The 3D-FICS will be used in experimental studies on small animal organs from which we will present the first obtained images.
Paul R. Bloemen, Iwan J. Dobbe, Coen D. Dijkman, Niels H. Kind, Maurice C. Aalders, Ton G. van Leeuwen, and Maria Siebes, "3D Fluorescence Imaging cryomicrotome system for multispectral structural, functional and molecular imaging of whole organs (Conference Presentation)," Proc. SPIE 10487, Multimodal Biomedical Imaging XIII, 104870B (Presented at SPIE BiOS: January 27, 2018; Published: 14 March 2018); https://doi.org/10.1117/12.2290142.5751463282001.
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