Towards standardised sample preparation in X- ray phase contrast imaging: comparison of eight protocols on equine superficial digital flexor tendon (SDFT)

X-ray phase contrast imaging (XPCi) is becoming increasingly important with regards to three-dimensional imaging of biological tissues, however research has largely been focused on technology development and exploring pre-clinical and research applications, with little consideration to optimal sample preparation techniques and their effect on image quality. This paper aims to fill this gap by comparing 8 different sample preparation techniques on ex vivo biological tissues. Equine superficial digital flexor tendons were prepared using various combinations of PBS, 10% neutral buffered formalin, 70% and 100% ethanol and imaged using an edge illumination XPCi system in a 3D printed container. Results demonstrated that chemical dehydration via ethanol is a recommended precursor to XPCi contrast generation with 100% ethanol the optimal dehydration agent for maximising contrast.