Medical micro ct systems

Intact knee joint from different animals were isolated, fixed overnight in 10% neutral buffered formalin (HT501128, Sigma) followed by washing with phosphate buffer saline (PBS) three times and transfer to 70% ethanol (v/v). Bones were then scanned using Scanco Medical micro-CT systems (Scanco, Wayne, PA, USA) at the core facility at the Musculoskeletal Center at Washington University in St. Louis (St. Louis, MO). Briefly, images were scanned at a resolution of 20 µm, slice increment 20 µm, voltage 55 kV, current 145 µA and exposure time of 200 ms. After scanning the knee joint area, three-dimensional images were constructed. X-ray analysis of isolated knee joints were performed using Faxitron UltraFocus 100 on automatic settings and at x5 magnification.

Intact femur and tibia from WT mice and NM-cKO animals were isolated, fixed overnight in 10% formalin followed by washing with Phosphate Buffer Saline (PBS) 3 times and transferred to 70% ethanol (v/v). Bones were then scanned using Scanco Medical micro-CT systems (Scanco, Wayne, PA, USA) at the core facility at the musculoskeletal center at Washington University (St. Louis, MO). Briefly, Images were scanned at a resolution of 20 μm, slice increment 20 μm, voltage 55 kV, current 145 μA and exposure time of 200 ms. After scanning the whole bone, contour was drawn from the growth plate toward trabecular and cortical regions of tibia of approximately 200 slices were analyzed for WT and NM-cKO bones, and 3D images were constructed.

Intact knee joints were harvested, fixed overnight in 10% neutral buffered formalin (HT501128, Sigma) then washing with Phosphate Buffer Saline (PBS) three times and transfered to 70% ethanol (v/v). Bones were scanned at a resolution of 20 µm, slice increment 10 µm, voltage 55 kV, current 145 µA and exposure time of 200 ms and 3D images were constructed using Scanco Medical micro-CT systems (Scanco, Wayne, PA, USA) at Washington University in St. Louis (St. Louis, MO).