Optical Projection Tomography with a Tissue Clearing Agent for Developmental and Reproductive Toxicology Studies.

Abstract

Developmental and reproductive toxicology (DART) testing represents an expensive and time-consuming stage in determining the toxicological profile of new chemical entities. Within DART studies, morphological evaluation of fetal skeletons for developmental abnormalities typically requires 7 to 14 days. Current processing techniques involve digestion of soft tissue using a strong base (KOH), followed by qualitative assessment of the remaining skeletal tissue by a fetal morphologist. Micro-computed tomography (micro-CT) has been proposed as a nondestructive image-based alternative for quantitative assessment of skeletal morphology. Such methods eliminate the need for extensive tissue processing and can be paired with quantitative analysis algorithms. However, due to the significant capital and operational expenses required for micro-CT imaging, this approach has yet to gain widespread traction and regulatory acceptance. A novel tissue clearing agent was used in 1-week-old rats to render soft tissue optically transparent. Alizarin red was used to stain the skeleton. High dynamic range optical trans-illumination images were then acquired with an optical-CT imaging system and rendered as three-dimensional images of skeletal structure. High dynamic range-based optical-CT imaging of chemically cleared tissues can rapidly generate high resolution (50-250 μm) reconstructions of whole skeletons. In summary, this study demonstrates that the combination of tissue clearing, optical imaging, and novel reconstruction algorithms may present a new paradigm for high-throughput evaluation of tissues in DART testing. Birth Defects Research 110:12-16, 2018. © 2017 Wiley Periodicals, Inc.