Development of heavy metal stain for Phase-contrast synchrotron X-Ray CT of in vitro nanofibrous heart valve scaffolds

In our previous works we successfully utilised the high-resolution in-line phase contrast capabilities of the I13-2 X-ray beamline to characterise the spatial distribution of cells, fibres and the mode of clustering, in a tissue-engineered heart valve made of jet-sprayed poly(ε-caprolactone) (PCL) nanofibrous material. However, image segmentation remains a challenge because both cells and PCL have indistinguishable X-ray attenuation. To overcome this challenge, we propose via this project developing a protocol to enhance the contrast of X-ray attenuation of the cells by using a heavy metal-based cell-specific stain, to allow for straightforward separation of cells and PCL by thresholding. This approach will streamline image acquisition and enhance biological data extraction, facilitating the wider adoption of the X-ray CT technique in studying the micro- and nano-structure of bioprosthetic heart valves, and the wider biomedical devices/implants.

Project Reference: MG40436
Estimated total in kind value £37,155