Visualization of color anatomy and molecular fluorescence in whole-mouse cryo-imaging

Gargesha, Madhusudhana; Qutaish, Mohammed Q.; Roy, Debashish; Steyer, Grant J.; Watanabe, Michiko; Wilson, David L.

doi:10.1016/j.compmedimag.2010.10.003

Cited by 17 publications

(14 citation statements)

References 17 publications

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“…We developed interactive 3D visualization that enables one to examine biodistribution in a mouse and then zoom to particular regions to examine densities or to zoom even further and see individual cells. All of this can be done within the context of volume rendered color brightfield data giving full anatomical context using specialized rendering reported earlier [12]. We encourage readers to examine movies in supplemental material.…”

Section: Discussionmentioning

confidence: 99%

Automatic Stem Cell Detection in Microscopic Whole Mouse Cryo-Imaging

Wuttisarnwattana

Gargesha

Hof

et al. 2016

IEEE Trans. Med. Imaging

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With its single cell sensitivity over volumes as large as or larger than a mouse, cryo-imaging enables imaging of stem cell biodistribution, homing, engraftment, and molecular mechanisms. We developed and evaluated a highly automated software tool to detect fluorescently labeled stem cells within very large (~200GB) cryo-imaging datasets. Cell detection steps are: preprocess, remove immaterial regions, spatially filter to create features, identify candidate pixels, classify pixels using bagging decision trees, segment cell patches, and perform 3D labeling. There are options for analysis and visualization. To train the classifier, we created synthetic images by placing realistic digital cell models onto cryo-images of control mice devoid of cells. Very good cell detection results were (precision=98.49%, recall=99.97%) for synthetic cryo-images, (precision=97.81%, recall=97.71%) for manually evaluated, actual cryo-images, and <1% false positives in control mice. An α-multiplier applied to features allows one to correct for experimental variations in cell brightness due to labeling. On dim cells (37% of standard brightness), with correction, we improved recall (49.26%→99.36%) without a significant drop in precision (99.99%→99.75%). With tail vein injection, multipotent adult progenitor cells in a graft-versus-host-disease model in the first days post injection were predominantly found in lung, liver, spleen, and bone marrow. Distribution was not simply related to blood flow. The lung contained clusters of cells while other tissues contained single cells. Our methods provided stem cell distribution anywhere in mouse with single cell sensitivity. Methods should provide a rational means of evaluating dosing, delivery methods, cell enhancements, and mechanisms for therapeutic cells.

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Section: Discussionmentioning

confidence: 99%

Automatic Stem Cell Detection in Microscopic Whole Mouse Cryo-Imaging

Wuttisarnwattana

Gargesha

Hof

et al. 2016

IEEE Trans. Med. Imaging

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“…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]. Following sacrifice, the mice were embedded in optimal cutting temperature (OCT) embedding compound and then snap frozen in liquid nitrogen.…”

Section: Cryo-imaging and Analysis Softwarementioning

confidence: 99%

“…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.…”

Section: Introductionmentioning

confidence: 99%

Novel T lymphocyte proliferation assessment using whole mouse cryo-imaging

Wuttisarnwattana

Raza

Eid³

et al. 2014

SPIE Proceedings

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New imaging technologies enable one to assess T-cell proliferation, an important feature of the immunological response. However, none of the traditional imaging modalities allow one to examine quantiatively T-cell function with microscopic resolution and single cell sensitivity over an entire mouse. To address this need, we established T-cells proliferation assays using 3D microscopic cryo-imaging. Assays include: (1) biodistribution of T-cells, (2) secondary lymphoid organ (SLO) volume measurement, (3) carboxyfluorescein succinimidyl ester (CFSE) dilution per cell as cells divide. To demonstrate the application, a graft-versus-host-disease (GVHD) model was used. 3D visualization show that T-cells specifically homed to the SLOs (spleen and lymph nodes) as well as GVHD target organs (such as GI-tract, liver, skin and thymus).The spleen was chosen as representative of the SLOs. For spleen size analysis, volumes of red and white pulp were measured. Spleen volumes of the allogeneic mice (with GVHD) were significantly larger than those of the syngeneic mice (without GVHD) at 72 to 120 hours post-transplant. For CFSE dilution approach, we employed color-coded volume rendering and probability density function (PDF) of single cell intensity to assess T-cell proliferation in the spleen. As compared to syngeneic T-cells, the allogeneic T-cells quickly aggregated in the spleen as indicated by increasing of CFSE signal over the first 48 hours. Then they rapidly proliferated as evidenced by reduced CFSE intensity (at 48-96 hours). Results suggest that assays can be used to study GVHD treatments using T-cell proliferation and biodistibution as assays. In summary, this is the first time that we are able to track and visualize T-cells in whole mouse with single cell sensitivity. We believe that our technique can be an alternative choice to traditional in vitro immunological proliferation assays by providing assessment of proliferation in an in vivo model.

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“…Inert, biocompatible, and multivalent plant viruses such as the cowpea mosaic virus (CPMV) have also shown to accumulate preferentially in the periphery of the vascular lumen and have been used in confocal and intravital imaging studies of neo-angiogenesis of even the smallest of capillaries [121]. Further, in situ three-dimensional fluorescence cryogenic imaging (3D cryo-imaging [122–124]) that allows for whole-body imaging has also been used for imaging/detection of small metastatic tumors (less than 0.5 mm) and associated stroma/endothelium following intravenous injection of fluorescent peptides that target the upregulated fibrin-fibronectin complexes [125]. …”

Section: Molecular Imaging Applicationsmentioning

confidence: 99%

Vascular targeting of nanoparticles for molecular imaging of diseased endothelium

Atukorale

Covarrubias

Bauer

et al. 2017

Advanced Drug Delivery Reviews

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This review seeks to highlight the enormous potential of targeted nanoparticles for molecular imaging applications. Being the closest point-of-contact, circulating nanoparticles can gain direct access to targetable molecular markers of disease that appear on the endothelium. Further, nanoparticles are ideally suitable to vascular targeting due to geometrically enhanced multivalent attachment on the vascular target. This natural synergy between nanoparticles, vascular targeting and molecular imaging can provide new avenues for diagnosis and prognosis of disease with quantitative precision. In addition to the obvious applications of targeting molecular signatures of vascular diseases (e.g., atherosclerosis), deep-tissue diseases often manifest themselves by continuously altering and remodeling their neighboring blood vessels (e.g., cancer). Thus, the remodeled endothelium provides a wide range of targets for nanoparticles and molecular imaging. To demonstrate the potential of molecular imaging, we present a variety of nanoparticles designed for molecular imaging of cancer or atherosclerosis using different imaging modalities.

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Visualization of color anatomy and molecular fluorescence in whole-mouse cryo-imaging

Cited by 17 publications

References 17 publications

Automatic Stem Cell Detection in Microscopic Whole Mouse Cryo-Imaging

Automatic Stem Cell Detection in Microscopic Whole Mouse Cryo-Imaging

Novel T lymphocyte proliferation assessment using whole mouse cryo-imaging

Vascular targeting of nanoparticles for molecular imaging of diseased endothelium

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