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Use of Micro X-ray Computed Tomography with Phosphotungstic Acid Preparation to Visualize Human Fibromuscular Tissue

 Jehoon O  ;  Hyun-Jin Kwon  ;  Shin Hyung Kim  ;  Tae-Hyeon Cho  ;  Hun-Mu Yang 
 Journal of Visualized Experiments : JoVE, Vol.151 : e59752, 2019 
Journal Title
 Journal of Visualized Experiments : JoVE 
Issue Date
Manual dissection and histological observation are common methods used to investigate human tissues. However, manual dissection can damage delicate structures while processing and histological observation provide limited information through cross-sectional imaging. Micro X-ray computed tomography (microCT) is an effective tool for obtaining three-dimensional information without damaging specimens. However, it shows limited efficiency in differentiating soft tissue parts. Use of contrast-enhancing agents, like phosphotungstic acid (PTA), can solve this problem by improving soft tissue contrast. We implemented microCT with PTA to investigate the human orbicularis retaining ligament (ORL), which is a delicate structure in the orbit area. In this method, harvested specimens are fixed in formalin, dehydrated in serial ethanol solutions, and stained with a PTA solution. After staining, microCT scanning, 3D reconstruction, and analysis are performed. Skin, ligaments, and muscles can be clearly visualized using this method. The specimen size and duration of staining are essential features of the method. The suitable specimen thickness was about 5-7 mm, above which the process was slowed, and the optimum duration was 5-7 days, below which an empty hole in the central area occasionally occurred. To maintain the location and direction of small pieces during cutting, sewing on the same region of each part is recommended. Furthermore, preliminary analyses of the anatomical structure are needed to correctly identify each piece. Parafilm can be used to prevent drying, but care should be taken to prevent specimen distortion. Our multidirectional observation showed that the ORL is composed of a multilayered meshwork of continuous plates, rather than thread-like fibers, as reported previously. These results suggest that microCT scanning with PTA is useful for examining specific compartments within complex structures of human tissue. It may be helpful in the analyses of cancer tissues, nerve tissues, and various organs, like the heart and liver.
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1. College of Medicine (의과대학) > Dept. of Anesthesiology and Pain Medicine (마취통증의학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Anatomy (해부학교실) > 1. Journal Papers
Yonsei Authors
Kim, Shin Hyung(김신형) ORCID logo https://orcid.org/0000-0003-4058-7697
Yang, Hun Mu(양헌무) ORCID logo https://orcid.org/0000-0003-1954-0114
O, Jehoon(오제훈) ORCID logo https://orcid.org/0000-0003-4351-3183
Cho, Tae-Hyeon(조태현) ORCID logo https://orcid.org/0000-0001-7230-620X
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