Laboratory-based x-ray phase contrast microscopy system for targeting in unstained soft-tissue samples

We propose an imaging system and methodology for mapping soft-tissue samples in three dimensions, with micron-scale and isotropic spatial resolution, with low-concentrations as well as in the absence of heavy metal staining. We used hard x-ray phase-contrast imaging for the x-ray ability to nondestructively probe the internal structure of opaque specimens and for enhanced contrast obtained by exploiting phase effects, even in cases with reduced or absent staining agents. To demonstrate its applicability to soft-tissue specimens, we built a compact system that is easily deployable in a laboratory setting. The imaging system is based on a conventional rotating anode x-ray tube and a state-of-the-art custom-made radiation detector. The systems performance is quantitatively assessed on a calibration standard. Its potential for soft-tissue microscopy is demonstrated on two biological specimens and benchmarked against gold-standard synchrotron data. We believe that the approach proposed here can be valuable as a bridging imaging modality for intravital correlative light and electron microscopy and be applied across disciplines where the three-dimensional morphology of pristine-condition soft tissues is a key element of the investigation.