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Talbot-defocus multiscan tomography using the synchrotron X-ray microscope to study the lacuno-canalicular network in mouse bone.

Nango N, Kubota S, Takeuchi A, Suzuki Y, Yashiro W, Momose A, Matsuo K - Biomed Opt Express (2013)

Bottom Line: The three-dimensional network of lacunae and canaliculi that regulates metabolism in bone contains osteocytes and their dendritic processes.We constructed a synchrotron radiation X-ray microscope for sequential tomography of mouse tibia first by using a Talbot interferometer to detect the degree of bone mineralization and then by using absorption contrast under a slightly defocused setting to enhance outline contrast thereby visualizing structures of the osteocyte lacuno-canalicular network.Thus, multiscan microscopic X-ray tomography is a powerful tool for analyzing bone mineralization in relation to the lacuno-canalicular network at the submicron resolution level.

View Article: PubMed Central - PubMed

Affiliation: Ratoc System Engineering Co., Ltd, 1-24-8 Sekiguchi, Bunkyo-ku, Tokyo 112-0014, Japan.

ABSTRACT
The three-dimensional network of lacunae and canaliculi that regulates metabolism in bone contains osteocytes and their dendritic processes. We constructed a synchrotron radiation X-ray microscope for sequential tomography of mouse tibia first by using a Talbot interferometer to detect the degree of bone mineralization and then by using absorption contrast under a slightly defocused setting to enhance outline contrast thereby visualizing structures of the osteocyte lacuno-canalicular network. The resultant pair of tomograms was precisely aligned with each other, allowing evaluation of mineral density in the vicinity of each osteocyte lacuna and canaliculus over the entire thickness of the cortical bone. Thus, multiscan microscopic X-ray tomography is a powerful tool for analyzing bone mineralization in relation to the lacuno-canalicular network at the submicron resolution level.

No MeSH data available.


Preparation of mouse bone sample. (a) Schematic presentation of tibia. The cortical bone sample used for imaging is represented in green. (b) The sample (green arrow) attached to the holder using double-sided adhesive tape (arrowhead). Scale bar, 1 mm.
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g002: Preparation of mouse bone sample. (a) Schematic presentation of tibia. The cortical bone sample used for imaging is represented in green. (b) The sample (green arrow) attached to the holder using double-sided adhesive tape (arrowhead). Scale bar, 1 mm.

Mentions: Tibias were isolated from euthanized 12 to 16 week-old female C57BL/6J mice (Clea, Tokyo, Japan). After removing muscles and soft tissues, tibias were fixed in 70% ethanol, infiltrated for three days in LR white resin (Electron Microscopy Sciences, Fort Washington, PA, USA) and embedded in the same resin. After waiting a day for specimens to solidify, the shaft of the long bone (diaphysis) was cut as a piece approximately 3 mm long and the posterior cortical bone was vertically sliced twice to make a 300 μm or less wide column with intact periosteum and endosteum surfaces (Fig. 2(a)Fig. 2


Talbot-defocus multiscan tomography using the synchrotron X-ray microscope to study the lacuno-canalicular network in mouse bone.

Nango N, Kubota S, Takeuchi A, Suzuki Y, Yashiro W, Momose A, Matsuo K - Biomed Opt Express (2013)

Preparation of mouse bone sample. (a) Schematic presentation of tibia. The cortical bone sample used for imaging is represented in green. (b) The sample (green arrow) attached to the holder using double-sided adhesive tape (arrowhead). Scale bar, 1 mm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3675870&req=5

g002: Preparation of mouse bone sample. (a) Schematic presentation of tibia. The cortical bone sample used for imaging is represented in green. (b) The sample (green arrow) attached to the holder using double-sided adhesive tape (arrowhead). Scale bar, 1 mm.
Mentions: Tibias were isolated from euthanized 12 to 16 week-old female C57BL/6J mice (Clea, Tokyo, Japan). After removing muscles and soft tissues, tibias were fixed in 70% ethanol, infiltrated for three days in LR white resin (Electron Microscopy Sciences, Fort Washington, PA, USA) and embedded in the same resin. After waiting a day for specimens to solidify, the shaft of the long bone (diaphysis) was cut as a piece approximately 3 mm long and the posterior cortical bone was vertically sliced twice to make a 300 μm or less wide column with intact periosteum and endosteum surfaces (Fig. 2(a)Fig. 2

Bottom Line: The three-dimensional network of lacunae and canaliculi that regulates metabolism in bone contains osteocytes and their dendritic processes.We constructed a synchrotron radiation X-ray microscope for sequential tomography of mouse tibia first by using a Talbot interferometer to detect the degree of bone mineralization and then by using absorption contrast under a slightly defocused setting to enhance outline contrast thereby visualizing structures of the osteocyte lacuno-canalicular network.Thus, multiscan microscopic X-ray tomography is a powerful tool for analyzing bone mineralization in relation to the lacuno-canalicular network at the submicron resolution level.

View Article: PubMed Central - PubMed

Affiliation: Ratoc System Engineering Co., Ltd, 1-24-8 Sekiguchi, Bunkyo-ku, Tokyo 112-0014, Japan.

ABSTRACT
The three-dimensional network of lacunae and canaliculi that regulates metabolism in bone contains osteocytes and their dendritic processes. We constructed a synchrotron radiation X-ray microscope for sequential tomography of mouse tibia first by using a Talbot interferometer to detect the degree of bone mineralization and then by using absorption contrast under a slightly defocused setting to enhance outline contrast thereby visualizing structures of the osteocyte lacuno-canalicular network. The resultant pair of tomograms was precisely aligned with each other, allowing evaluation of mineral density in the vicinity of each osteocyte lacuna and canaliculus over the entire thickness of the cortical bone. Thus, multiscan microscopic X-ray tomography is a powerful tool for analyzing bone mineralization in relation to the lacuno-canalicular network at the submicron resolution level.

No MeSH data available.