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Proceedings Paper

Cryo-imaging in a toxicological study on mouse fetuses
Author(s): Debashish Roy; Madhusudhana Gargesha; Eddie Sloter; Michiko Watanabe; David Wilson
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Paper Abstract

We applied the Case cryo-imaging system to detect signals of developmental toxicity in transgenic mouse fetuses resulting from maternal exposure to a developmental environmental toxicant (2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD). We utilized a fluorescent transgenic mouse model that expresses Green Fluorescent Protein (GFP) exclusively in smooth muscles under the control of the smooth muscle gamma actin (SMGA) promoter (SMGA/EGFP mice kindly provided by J. Lessard, U. Cincinnati). Analysis of cryo-image data volumes, comprising of very high-resolution anatomical brightfield and molecular fluorescence block face images, revealed qualitative and quantitative morphological differences in control versus exposed fetuses. Fetuses randomly chosen from pregnant females euthanized on gestation day (GD) 18 were either manually examined or cryo-imaged. For cryo-imaging, fetuses were embedded, frozen and cryo-sectioned at 20 μm thickness and brightfield color and fluorescent block-face images were acquired with an in-plane resolution of ≈15 μm. Automated 3D volume visualization schemes segmented out the black embedding medium and blended fluorescence and brightfield data to produce 3D reconstructions of all fetuses. Comparison of Treatment groups TCDD GD13, TCDD GD14 and control through automated analysis tools highlighted differences not observable by prosectors performing traditional fresh dissection. For example, severe hydronephrosis, suggestive of irreversible kidney damage, was detected by cryoimaging in fetuses exposed to TCDD. Automated quantification of total fluorescence in smooth muscles revealed suppressed fluorescence in TCDD-exposed fetuses. This application demonstrated that cryo-imaging can be utilized as a routine high-throughput screening tool to assess the effects of potential toxins on the developmental biology of small animals.

Paper Details

Date Published: 9 March 2010
PDF: 9 pages
Proc. SPIE 7626, Medical Imaging 2010: Biomedical Applications in Molecular, Structural, and Functional Imaging, 76260X (9 March 2010);
Show Author Affiliations
Debashish Roy, Case Western Reserve Univ. (United States)
Madhusudhana Gargesha, Case Western Reserve Univ. (United States)
Eddie Sloter, WIL Research Labs., LLC (United States)
Michiko Watanabe, Case Western Reserve Univ. School of Medicine (United States)
David Wilson, Case Western Reserve Univ. (United States)
Univ. Hospitals of Cleveland (United States)

Published in SPIE Proceedings Vol. 7626:
Medical Imaging 2010: Biomedical Applications in Molecular, Structural, and Functional Imaging
Robert C. Molthen; John B. Weaver, Editor(s)

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