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Histological evaluations using <t>ALP</t> and <t>ARS</t> <t>staining</t> for cultured cells on fabricated ceramics. ( a ) ALP staining for cultured specimens using a bone differentiation induction medium for 1–2 weeks after preculture. ARS staining ( b ) without and ( c ) with a bone differentiation induction medium for 1–2 weeks after preculture.
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Histological evaluations using <t>ALP</t> and <t>ARS</t> <t>staining</t> for cultured cells on fabricated ceramics. ( a ) ALP staining for cultured specimens using a bone differentiation induction medium for 1–2 weeks after preculture. ARS staining ( b ) without and ( c ) with a bone differentiation induction medium for 1–2 weeks after preculture.
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Histological evaluations using <t>ALP</t> and <t>ARS</t> <t>staining</t> for cultured cells on fabricated ceramics. ( a ) ALP staining for cultured specimens using a bone differentiation induction medium for 1–2 weeks after preculture. ARS staining ( b ) without and ( c ) with a bone differentiation induction medium for 1–2 weeks after preculture.
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Histological evaluations using <t>ALP</t> and <t>ARS</t> <t>staining</t> for cultured cells on fabricated ceramics. ( a ) ALP staining for cultured specimens using a bone differentiation induction medium for 1–2 weeks after preculture. ARS staining ( b ) without and ( c ) with a bone differentiation induction medium for 1–2 weeks after preculture.
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FIGURE 4 Thunnus orientalis, distribution of osteoblasts and osteoclasts in vertebrae. (a) Double staining of <t>ALP</t> and TRAP of a juvenile vertebral body. The total length of the juvenile of (a) is 11.2 cm. (b–d) Magnified images of (b) the lateral ridge, (c) the base of the dorsal arch, and (d) the base of the hemal arch base marked with rectangles in (a). Arrows and arrowheads indicate ALP‐positive osteoblasts and TRAP‐positive osteoclasts, respectively. (e) HE and alcian blue staining of a lateral ridge of an adult vertebra. (f, g) Magnified images of (f) the distal edge and (g) the surface of the lateral ridge marked with rectangles in (e). (f) Black‐filled arrowheads indicate osteoblasts accumulating on the edge of the ridge. Asterisks indicate the small cavities in which osteoblasts are embedded. Branches from black circles indicate osteocyte lacunae. The cavities are much larger than the osteocyte lacunae. (g) The arrowhead indicates osteoblasts lined up along the surface of the ridge. No alcian‐ blue‐positive cartilage was observed. Scale bars: (a–d) 100 µm, (e) 500 µm, and (f, g) 50 µm. ALP, alkaline <t>phosphatase;</t> bv, blood vessel; HE, hematoxylin and eosin; sc, spinal cord; TRAP, tartrate‐resistant acid phosphatase.
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FIGURE 4 Thunnus orientalis, distribution of osteoblasts and osteoclasts in vertebrae. (a) Double staining of <t>ALP</t> and TRAP of a juvenile vertebral body. The total length of the juvenile of (a) is 11.2 cm. (b–d) Magnified images of (b) the lateral ridge, (c) the base of the dorsal arch, and (d) the base of the hemal arch base marked with rectangles in (a). Arrows and arrowheads indicate ALP‐positive osteoblasts and TRAP‐positive osteoclasts, respectively. (e) HE and alcian blue staining of a lateral ridge of an adult vertebra. (f, g) Magnified images of (f) the distal edge and (g) the surface of the lateral ridge marked with rectangles in (e). (f) Black‐filled arrowheads indicate osteoblasts accumulating on the edge of the ridge. Asterisks indicate the small cavities in which osteoblasts are embedded. Branches from black circles indicate osteocyte lacunae. The cavities are much larger than the osteocyte lacunae. (g) The arrowhead indicates osteoblasts lined up along the surface of the ridge. No alcian‐ blue‐positive cartilage was observed. Scale bars: (a–d) 100 µm, (e) 500 µm, and (f, g) 50 µm. ALP, alkaline <t>phosphatase;</t> bv, blood vessel; HE, hematoxylin and eosin; sc, spinal cord; TRAP, tartrate‐resistant acid phosphatase.
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Histological evaluations using ALP and ARS staining for cultured cells on fabricated ceramics. ( a ) ALP staining for cultured specimens using a bone differentiation induction medium for 1–2 weeks after preculture. ARS staining ( b ) without and ( c ) with a bone differentiation induction medium for 1–2 weeks after preculture.

Journal: International Journal of Molecular Sciences

Article Title: Preferred Orientation of Hydroxyapatite Ceramics Along the c -Axis Promotes Osteoblast Differentiation

doi: 10.3390/ijms252312926

Figure Lengend Snippet: Histological evaluations using ALP and ARS staining for cultured cells on fabricated ceramics. ( a ) ALP staining for cultured specimens using a bone differentiation induction medium for 1–2 weeks after preculture. ARS staining ( b ) without and ( c ) with a bone differentiation induction medium for 1–2 weeks after preculture.

Article Snippet: For ALP staining, the cells were incubated with ALP substrate solution (Wako Pure Chemical Industries, Osaka, Japan) in an incubator at 37 °C for 30 min. Then, the cells were washed with sterile water and observed.

Techniques: Staining, Cell Culture

FIGURE 4 Thunnus orientalis, distribution of osteoblasts and osteoclasts in vertebrae. (a) Double staining of ALP and TRAP of a juvenile vertebral body. The total length of the juvenile of (a) is 11.2 cm. (b–d) Magnified images of (b) the lateral ridge, (c) the base of the dorsal arch, and (d) the base of the hemal arch base marked with rectangles in (a). Arrows and arrowheads indicate ALP‐positive osteoblasts and TRAP‐positive osteoclasts, respectively. (e) HE and alcian blue staining of a lateral ridge of an adult vertebra. (f, g) Magnified images of (f) the distal edge and (g) the surface of the lateral ridge marked with rectangles in (e). (f) Black‐filled arrowheads indicate osteoblasts accumulating on the edge of the ridge. Asterisks indicate the small cavities in which osteoblasts are embedded. Branches from black circles indicate osteocyte lacunae. The cavities are much larger than the osteocyte lacunae. (g) The arrowhead indicates osteoblasts lined up along the surface of the ridge. No alcian‐ blue‐positive cartilage was observed. Scale bars: (a–d) 100 µm, (e) 500 µm, and (f, g) 50 µm. ALP, alkaline phosphatase; bv, blood vessel; HE, hematoxylin and eosin; sc, spinal cord; TRAP, tartrate‐resistant acid phosphatase.

Journal: Journal of morphology

Article Title: Lateral bone ridge expansion and internal tissue replacement for vertebral body growth in Pacific bluefin tuna Thunnus orientalis.

doi: 10.1002/jmor.21666

Figure Lengend Snippet: FIGURE 4 Thunnus orientalis, distribution of osteoblasts and osteoclasts in vertebrae. (a) Double staining of ALP and TRAP of a juvenile vertebral body. The total length of the juvenile of (a) is 11.2 cm. (b–d) Magnified images of (b) the lateral ridge, (c) the base of the dorsal arch, and (d) the base of the hemal arch base marked with rectangles in (a). Arrows and arrowheads indicate ALP‐positive osteoblasts and TRAP‐positive osteoclasts, respectively. (e) HE and alcian blue staining of a lateral ridge of an adult vertebra. (f, g) Magnified images of (f) the distal edge and (g) the surface of the lateral ridge marked with rectangles in (e). (f) Black‐filled arrowheads indicate osteoblasts accumulating on the edge of the ridge. Asterisks indicate the small cavities in which osteoblasts are embedded. Branches from black circles indicate osteocyte lacunae. The cavities are much larger than the osteocyte lacunae. (g) The arrowhead indicates osteoblasts lined up along the surface of the ridge. No alcian‐ blue‐positive cartilage was observed. Scale bars: (a–d) 100 µm, (e) 500 µm, and (f, g) 50 µm. ALP, alkaline phosphatase; bv, blood vessel; HE, hematoxylin and eosin; sc, spinal cord; TRAP, tartrate‐resistant acid phosphatase.

Article Snippet: To detect osteoblasts, we mounted 100 μL alkaline phosphatase (ALP) substrate solution (Takara) and incubated the sections for 20min at 37°C.

Techniques: Double Staining, Staining

FIGURE 5 Thunnus orientalis, osteoblasts in the small cavities of vertebrae. (a) Schematic illustration showing the positions of the slices (b–d) in vertebrae. (b–d) Double staining of ALP and TRAP of the vertebrae of the juveniles. Green lines in (b–d) indicate the outline of the bones, which correspond to that of (a). Arrows indicate the direction of osteoblast alignment. (b) The lateral part of an hourglass‐shaped bone from which the lateral ridge extends. Osteoblasts are embedded in the small cavities, and the direction of osteoblasts is circular in the hourglass‐shaped parts and lateral in the lateral ridge. No osteoclasts are observed in the small cavities. Arrowheads indicate osteoclasts forming bone marrow cavities. (c) The distal end of the lateral ridge. Osteoblasts are laterally aligned in the distal region of the lateral ridge, and the alignments are nearly parallel. (d) Neural arch. Osteoblasts are arranged radially toward the dorsal side. The total length of juveniles of (b, c) is 39 cm, and (d) is 20 cm. Scale bars: 500 µm. ALP, alkaline phosphatase; sc, spinal cord; TRAP, tartrate‐resistant acid phosphatase.

Journal: Journal of morphology

Article Title: Lateral bone ridge expansion and internal tissue replacement for vertebral body growth in Pacific bluefin tuna Thunnus orientalis.

doi: 10.1002/jmor.21666

Figure Lengend Snippet: FIGURE 5 Thunnus orientalis, osteoblasts in the small cavities of vertebrae. (a) Schematic illustration showing the positions of the slices (b–d) in vertebrae. (b–d) Double staining of ALP and TRAP of the vertebrae of the juveniles. Green lines in (b–d) indicate the outline of the bones, which correspond to that of (a). Arrows indicate the direction of osteoblast alignment. (b) The lateral part of an hourglass‐shaped bone from which the lateral ridge extends. Osteoblasts are embedded in the small cavities, and the direction of osteoblasts is circular in the hourglass‐shaped parts and lateral in the lateral ridge. No osteoclasts are observed in the small cavities. Arrowheads indicate osteoclasts forming bone marrow cavities. (c) The distal end of the lateral ridge. Osteoblasts are laterally aligned in the distal region of the lateral ridge, and the alignments are nearly parallel. (d) Neural arch. Osteoblasts are arranged radially toward the dorsal side. The total length of juveniles of (b, c) is 39 cm, and (d) is 20 cm. Scale bars: 500 µm. ALP, alkaline phosphatase; sc, spinal cord; TRAP, tartrate‐resistant acid phosphatase.

Article Snippet: To detect osteoblasts, we mounted 100 μL alkaline phosphatase (ALP) substrate solution (Takara) and incubated the sections for 20min at 37°C.

Techniques: Double Staining