Abstract
High-contrast, high-resolution imaging of biomedical specimens is indispensable for studying organ function and pathologies. Conventional histology, the gold standard for soft-tissue visualization, is limited by its anisotropic spatial resolution, elaborate sample preparation, and lack of quantitative image information. X-ray absorption or phase tomography have been identified as promising alternatives enabling non-destructive, distortion-free three-dimensional (3D) imaging. However, reaching sufficient contrast and resolution with a simple experimental procedure remains a major challenge. Here, we present a solution based on x-ray phase tomography through speckle-based imaging (SBI). We demonstrate on a mouse kidney that SBI delivers comprehensive 3D maps of hydrated, unstained soft tissue, revealing its microstructure and delivering quantitative tissue-density values at a density resolution of better than 2 m g / c m 3 and spatial resolution of better than 8 µm. We expect that SBI virtual histology will find widespread application in biomedicine and will open up new possibilities for research and histopathology.
Highlights
The microscopic visualization of human and animal tissue is a fundamental part of pathological investigations in research and clinical practice
As the phase tomogram can be virtually sectioned along any desired plane, the correlation with slices obtained by conventional histology, which was performed on the same specimen after x-ray imaging, is straightforward
The microstructural detail of the hydrated kidney tissue is clearly visualized in the phase volume slices and matches that seen by conventional histology with hemotoxylin and eosin (H&E) stain, see Figs. 2(b) and 2(f )–2(h)
Summary
The microscopic visualization of human and animal tissue is a fundamental part of pathological investigations in research and clinical practice. Softtissue discrimination requires the use of contrast agents, and the visualization of blood vessels is commonly achieved by corrosion casting with a resin [3,4,5,6,7,8,9] These approaches, bear the risk of altering the tissue structure, are susceptible to incomplete tissue or vessel penetration, and are often incompatible with complementary follow-up imaging techniques including conventional histology. The x-ray phase-contrast imaging methods employed for this purpose to date, such as propagation-based imaging (PBI) [16,17,18] and x-ray grating interferometry (XGI) [19,20,21], have various limitations that have prevented their practical use and wider uptake for virtual histology These include an elaborate experimental setup with costly, absorbing x-ray optical elements and insufficient spatial resolution for XGI and constraints on the size and nature of the sample and non-quantitative results for PBI. We demonstrate the unexplored potential of SBI x-ray phase tomography for the application of 3D virtual histology on a whole unstained, hydrated kidney of a healthy mouse imaged at a synchrotron radiation facility
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