3D Cryo‐Imaging: A Very High‐Resolution View of the Whole Mouse

Abstract

We developed the Case Cryo-imaging system that provides information rich, very high-resolution, color brightfield, and molecular fluorescence images of a whole mouse using a section-and-image block-face imaging technology. The system consists of a mouse-sized, motorized cryo-microtome with special features for imaging, a modified, brightfield/ fluorescence microscope, and a robotic xyz imaging system positioner, all of which is fully automated by a control system. Using the robotic system, we acquired microscopic tiled images at a pixel size of 15.6 mm over the block face of a whole mouse sectioned at 40 mm, with a total data volume of 55 GB. Viewing 2D images at multiple resolutions, we identified small structures such as cardiac vessels, muscle layers, villi of the small intestine, the optic nerve, and layers of the eye. Cryo-imaging was also suitable for imaging embryo mutants in 3D. A mouse, in which enhanced green fluorescent protein was expressed under gamma actin promoter in smooth muscle cells, gave clear 3D views of smooth muscle in the urogenital and gastrointestinal tracts. With cryo-imaging, we could obtain 3D vasculature down to 10 mm, over very large regions of mouse brain. Software is fully automated with fully programmable imaging/sectioning protocols, email notifications, and automatic volume visualization. With a unique combination of field-of-view, depth of field, contrast, and resolution, the Case Cryo-imaging system fills the gap between whole animal in vivo imaging and histology.

Voxel-based plaque classification in coronary intravascular optical coherence tomography images using decision trees

“…Cryo-imaging [5] is a novel imaging technique where block face images of frozen tissue samples are captured with an episcopic microscope and a CCD camera. The method allows for cutting a thin section of the sample, acquiring a bright field image and a fluorescence image.…”

Novel T lymphocyte proliferation assessment using whole mouse cryo-imaging

“…Cryoimaging system provides 3-dimensional, tiled, microscopic anatomical bright field and molecular fluorescence image over entire mouse [9]. With sufficient sensitivity for detection of single cells, cryo-imaging permits unique quantitative analyses of fluorescently-labeled cells using specialized image analysis and visualization software [10,11].…”

“…To meet the missing requirement, we developed a novel lymphocyte proliferation assessment based on cryo-imaging. Cryo-imaging [9][10][11] is a fullyautomated, whole mouse section-and-image system which provides 3-dimensional, tiled, microscopic anatomical bright field and molecular fluorescence image over vast volumes. It provides single cell detection anywhere in mouse and determines cell densities far below that which can be observed with any other imaging technologies such as MRI, CT, PET, SPECT and BLI.…”

Model-Based Vasculature Extraction From Optical Fluorescence Cryomicrotome Images

“…Despite the requirement of a specialized set up of experimental microscopes or digital cameras, this potential means that the cryomicrotome modality is now an increasingly established imaging technique with several variations of the microtome set-up published in literature. Rosenthal et al [32] and Roy et al [33] implemented fluorescence and brightfield (Wilson et al [34], Roy et al [35], Steyer et al [36]) microtome sectioning, respectively. A similar methodology has been employed by LeGrice et al [37] to capture serial confocal microscope images over large tissue areas (Sands et al [38], Annese et al [39], Gerneke et al [40], Pope et al [41], Sand et al [42]).…”