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Graphical abstract of experimental design for imaging intravital C’Dot uptake and clearance in bone with 2-photon <t>microscopy.</t> A) Bone and osteocytes were exposed to fluorescent nanoparticles through a local subcutaneous injection above the third metatarsal (MT3) bone in a mouse hind paw. After C’Dot incubation, the MT3 was surgically isolated and secured for <t>multiphoton</t> imaging. B-D) Example images of C’Dots in the lacuno-canalicular network are shown. E-F) The high resolution of our imaging system enables subcellular localization. G) A short C’Dot incubation time and high frame averaging allows visualization of the canalicular network. H) Images can be analyzed for clearance kinetics of nanoparticle signal (from previous paper ) as well as subcellular distribution of signal. Scale bars = 10 μm.
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Graphical abstract of experimental design for imaging intravital C’Dot uptake and clearance in bone with 2-photon <t>microscopy.</t> A) Bone and osteocytes were exposed to fluorescent nanoparticles through a local subcutaneous injection above the third metatarsal (MT3) bone in a mouse hind paw. After C’Dot incubation, the MT3 was surgically isolated and secured for <t>multiphoton</t> imaging. B-D) Example images of C’Dots in the lacuno-canalicular network are shown. E-F) The high resolution of our imaging system enables subcellular localization. G) A short C’Dot incubation time and high frame averaging allows visualization of the canalicular network. H) Images can be analyzed for clearance kinetics of nanoparticle signal (from previous paper ) as well as subcellular distribution of signal. Scale bars = 10 μm.
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Graphical abstract of experimental design for imaging intravital C’Dot uptake and clearance in bone with 2-photon <t>microscopy.</t> A) Bone and osteocytes were exposed to fluorescent nanoparticles through a local subcutaneous injection above the third metatarsal (MT3) bone in a mouse hind paw. After C’Dot incubation, the MT3 was surgically isolated and secured for <t>multiphoton</t> imaging. B-D) Example images of C’Dots in the lacuno-canalicular network are shown. E-F) The high resolution of our imaging system enables subcellular localization. G) A short C’Dot incubation time and high frame averaging allows visualization of the canalicular network. H) Images can be analyzed for clearance kinetics of nanoparticle signal (from previous paper ) as well as subcellular distribution of signal. Scale bars = 10 μm.
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Graphical abstract of experimental design for imaging intravital C’Dot uptake and clearance in bone with 2-photon <t>microscopy.</t> A) Bone and osteocytes were exposed to fluorescent nanoparticles through a local subcutaneous injection above the third metatarsal (MT3) bone in a mouse hind paw. After C’Dot incubation, the MT3 was surgically isolated and secured for <t>multiphoton</t> imaging. B-D) Example images of C’Dots in the lacuno-canalicular network are shown. E-F) The high resolution of our imaging system enables subcellular localization. G) A short C’Dot incubation time and high frame averaging allows visualization of the canalicular network. H) Images can be analyzed for clearance kinetics of nanoparticle signal (from previous paper ) as well as subcellular distribution of signal. Scale bars = 10 μm.
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Graphical abstract of experimental design for imaging intravital C’Dot uptake and clearance in bone with 2-photon <t>microscopy.</t> A) Bone and osteocytes were exposed to fluorescent nanoparticles through a local subcutaneous injection above the third metatarsal (MT3) bone in a mouse hind paw. After C’Dot incubation, the MT3 was surgically isolated and secured for <t>multiphoton</t> imaging. B-D) Example images of C’Dots in the lacuno-canalicular network are shown. E-F) The high resolution of our imaging system enables subcellular localization. G) A short C’Dot incubation time and high frame averaging allows visualization of the canalicular network. H) Images can be analyzed for clearance kinetics of nanoparticle signal (from previous paper ) as well as subcellular distribution of signal. Scale bars = 10 μm.
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Graphical abstract of experimental design for imaging intravital C’Dot uptake and clearance in bone with 2-photon <t>microscopy.</t> A) Bone and osteocytes were exposed to fluorescent nanoparticles through a local subcutaneous injection above the third metatarsal (MT3) bone in a mouse hind paw. After C’Dot incubation, the MT3 was surgically isolated and secured for <t>multiphoton</t> imaging. B-D) Example images of C’Dots in the lacuno-canalicular network are shown. E-F) The high resolution of our imaging system enables subcellular localization. G) A short C’Dot incubation time and high frame averaging allows visualization of the canalicular network. H) Images can be analyzed for clearance kinetics of nanoparticle signal (from previous paper ) as well as subcellular distribution of signal. Scale bars = 10 μm.
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Graphical abstract of experimental design for imaging intravital C’Dot uptake and clearance in bone with 2-photon <t>microscopy.</t> A) Bone and osteocytes were exposed to fluorescent nanoparticles through a local subcutaneous injection above the third metatarsal (MT3) bone in a mouse hind paw. After C’Dot incubation, the MT3 was surgically isolated and secured for <t>multiphoton</t> imaging. B-D) Example images of C’Dots in the lacuno-canalicular network are shown. E-F) The high resolution of our imaging system enables subcellular localization. G) A short C’Dot incubation time and high frame averaging allows visualization of the canalicular network. H) Images can be analyzed for clearance kinetics of nanoparticle signal (from previous paper ) as well as subcellular distribution of signal. Scale bars = 10 μm.
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Graphical abstract of experimental design for imaging intravital C’Dot uptake and clearance in bone with 2-photon microscopy. A) Bone and osteocytes were exposed to fluorescent nanoparticles through a local subcutaneous injection above the third metatarsal (MT3) bone in a mouse hind paw. After C’Dot incubation, the MT3 was surgically isolated and secured for multiphoton imaging. B-D) Example images of C’Dots in the lacuno-canalicular network are shown. E-F) The high resolution of our imaging system enables subcellular localization. G) A short C’Dot incubation time and high frame averaging allows visualization of the canalicular network. H) Images can be analyzed for clearance kinetics of nanoparticle signal (from previous paper ) as well as subcellular distribution of signal. Scale bars = 10 μm.

Journal: Scientific Reports

Article Title: Real-time visualization and modulation of endocytic dynamics in osteocytes in vivo

doi: 10.1038/s41598-025-05735-1

Figure Lengend Snippet: Graphical abstract of experimental design for imaging intravital C’Dot uptake and clearance in bone with 2-photon microscopy. A) Bone and osteocytes were exposed to fluorescent nanoparticles through a local subcutaneous injection above the third metatarsal (MT3) bone in a mouse hind paw. After C’Dot incubation, the MT3 was surgically isolated and secured for multiphoton imaging. B-D) Example images of C’Dots in the lacuno-canalicular network are shown. E-F) The high resolution of our imaging system enables subcellular localization. G) A short C’Dot incubation time and high frame averaging allows visualization of the canalicular network. H) Images can be analyzed for clearance kinetics of nanoparticle signal (from previous paper ) as well as subcellular distribution of signal. Scale bars = 10 μm.

Article Snippet: Fluorescent signal inside the functionally isolated MT3 was visualized with multiphoton microscopy (MPM) (Bergamo II, Thorlabs) as previously described .

Techniques: Imaging, Microscopy, Injection, Incubation, Isolation