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This publication documents 3D image stacks from HR-pQCT imaging of a femur diaphysis, as well as image stacks for two in-situ loaded fracture mechanics specimens observed with 3D X-ray microscopy. Imaging For HR-qQCT: HR-pQCT scans were acquired by Rachel Surowiec using an XtremeCT II scanner (SCANCO Medical AG, Bruttisellen, Switzerland) within the Musculoskeletal Function, Imaging and Tissue (MSK-FIT) Resource Core of the Indiana Center for Musculoskeletal Health’s Clinical Research Center (Indiana University, Indianapolis, IN). Scans are performed at 60.7 um resolution, a 68 kV, 1467 uA, 43 ms integration time, 1 frame averaging. Raw scans are ‘.RSQ’ file types. The ISQ file type were read into ImageJ using the Import-KHKs Scanco uCT ISQ file reader plug-in, and exported as bmp image stacks, image stacks are provided in two parts. Reconstructed images are rotated in dataviewer so that all bones are in the same orientation (prox/distal/anterior/posterior for the Femur). For in-situ fracture mechanics experiments: 3D scans were acquired by Glynn Gallaway using a 3-point bending rig for single edged notched bend specimens with a Deben CT5000N load cell (Deben, Bury St. Edmunds, UK) in a Zeiss XRADIA 510 Versa 3D X-Ray microscope (Carl Zeiss AG, Baden-Württemberg, Germany) at Purdue University. The 3-point bending frame had a span 20 mm with X-ray transparent, glassy carbon supports. To maintain hydration, the beam was wrapped in a plastic film slit at the notch. Displacements were applied at 0.1 mm/min. Load cell outputs were monitored and recorded. Displacements are held constant during image acquisitions. The first 3D image was obtained at the onset of non-linearity. Subsequently, the displacement was increased until a load increase of 10 N was observed, and another image was obtained. This sequence was repeated 6-times until peak load. 3D X-ray images were acquired with a resolution of 4.5 um, exposure time 5 sec., 801 projections, 120 kV, 10 W, 4 x objective, and a LE2 filter. X-ray projections were processed through XRADIA Scout-and-Scan Reconstructor. A recursive Gaussian smoothing filter (s=1 pixel) was applied to reduce image artifacts. Image stacks are exported as tiff files and provided individually for each load step and specimen. Two experiments are documented (beam 1 and beam 2). MaterialstThe diaphysis of a human (92-year-old, male) cadaveric femur was obtained through the Indiana University School of Medicine Anatomical Donation Program.