J Vet Sci.  2011 Jun;12(2):107-113. 10.4142/jvs.2011.12.2.107.

Real time observation of mouse fetal skeleton using a high resolution X-ray synchrotron

• Chang DW
• Kim B
• Shin JH
• Yun YM
• Je JH
• Hwu YK
• Yoon JH
• Seong JK

• KMID: 1106551
• DOI: http://doi.org/10.4142/jvs.2011.12.2.107

Abstract

The X-ray synchrotron is quite different from conventional radiation sources. This technique may expand the capabilities of conventional radiology and be applied in novel manners for special cases. To evaluate the usefulness of X-ray synchrotron radiation systems for real time observations, mouse fetal skeleton development was monitored with a high resolution X-ray synchrotron. A non-monochromatized X-ray synchrotron (white beam, 5C1 beamline) was employed to observe the skeleton of mice under anesthesia at embryonic day (E)12, E14, E15, and E18. At the same time, conventional radiography and mammography were used to compare with X-ray synchrotron. After synchrotron radiation, each mouse was sacrificed and stained with Alizarin red S and Alcian blue to observe bony structures. Synchrotron radiation enabled us to view the mouse fetal skeleton beginning at gestation. Synchrotron radiation systems facilitate real time observations of the fetal skeleton with greater accuracy and magnification compared to mammography and conventional radiography. Our results show that X-ray synchrotron systems can be used to observe the fine structures of internal organs at high magnification.

Keyword

mouse; real-time observation; X-ray synchrotron

MeSH Terms

Animals
Bone and Bones/*anatomy & histology/radiography
Female
Fetus/*anatomy & histology/radiography
Histocytochemistry
Mice
Mice, Inbred ICR
Pregnancy
Synchrotrons
X-Rays

Figure

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  Fig. 1 Schematic diagram of the experimental set-up. Polychromatic X-rays (A), are emitted from the bending magnet device (b) of the storage ring (a) then pass through two slits (c: fixed one in the vacuum, d: changeable in the air) to control the beam size, attenuator set (e) for acquiring a good background image, and sample (f). The X-rays are processed by the scintillator (g) and the resulting image information is then converted into visible light (B). This visible light is magnified (C) by lens (i) after being reflected by the mirror (h) and transmitted to a computer or digital video recorder by the CCD camera (j).

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  Fig. 2 Photograph of a pregnant mouse suspended in the rectangular positioner with surgical ties. Synchrotron radiation imaging was performed with the mouse in an upright position.

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  Fig. 3 Photographs of a mouse fetus thorax obtained by synchrotron radiation at embryonic day (E) 14 (A), E15 (B), and E18 (C, D). At E18, fetal ribs (arrows) and thoracic vertebra (arrowhead) were observed. Fetal ribs were also visualized at E14 and E15 but with lower definition.

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  Fig. 4 Photographs of a fetus stained with Alizarin red S and Alcian blue (A) and images of fetal ribs obtained with synchrotron radiation (B), mammography (C), and conventional radiography (D) in a pregnant mouse at E14. Although the ribs had not yet calcified (arrow), they were visualized with synchrotron radiation (arrowhead). However, the ribs were not observed with the mammography equipment or conventional radiography.

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  Fig. 5 Photographs of a fetus stained with Alizarin red S and Alcian blue (A) and images of fetal ribs obtained with synchrotron radiation (B), mammography (C), and conventional radiography (D) of a pregnant mouse at E18. Ossified fetal ribs stained red were observed (arrow), and ribs were visualized by synchrotron radiation with high resolution (arrowhead). Fetuses (white arrows) were identified by mammography and conventional radiograph; however, the fetuses were too small to identify anatomic structures.

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  Fig. 6 Photographs of a fetus removed from a pregnant mouse at E18. The fetus was stained with Alizarin red S and Alcian blue. It was about 18 mm in length from crown to rump. Rectangles represent the field of views imaged by synchrotron radiation. A-I: comparisons of synchrotron radiation (1) and double-staining (2) images of the fetal mouse at E18, A: The fetal metatarsus showed calcified metatarsal bones (arrow), B: Distal femur (arrow), proximal tibia (white arrow), and fibula (arrowhead), C: Distal humerus (arrow), proximal radius (arrowhead), and ulna (white arrow), D: Scapula (arrow), proximal humerus (arrowhead), and spinous process of the scapula (white arrow), E: Mandible (arrow) and nasal bone (white arrow), F: Atlas (arrow) and foramen magnum (white arrow), G: Last rib (arrow) and first lumbar vertebra (white arrows), H: Body of ilium (arrow), sacrum (white arrow), I: Pubis (arrow), sacrum (white arrow) and ischium (arrowhead).

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