Extending two-dimensional histology into the third dimension through conventional micro computed tomography.
Histological examination achieves sub-micrometer resolution laterally. In the third dimension, however, resolution is limited to section thickness. In addition, histological sectioning and mounting sections on glass slides introduce tissue-dependent stress and strain. In contrast, state-of-the-art hard X-ray micro computed tomography (μCT) systems provide isotropic sub-micrometer resolution and avoid sectioning artefacts. The drawback of μCT in the absorption contrast mode for visualising physically soft tissue is a low attenuation difference between anatomical features. In this communication, we demonstrate that formalin-fixed paraffin-embedded human cerebellum yields appropriate absorption contrast in laboratory-based μCT data, comparable to conventional histological sections. Purkinje cells, for example, are readily visible. In order to investigate the pros and cons of complementary approaches, two- and three-dimensional data were manually and automatically registered. The joint histogram of histology and the related μCT slice allows for a detailed discussion on how to integrate two-dimensional information from histology into a three-dimensional tomography dataset. This methodology is not only rewarding for the analysis of the human cerebellum, but it also has relevance for investigations of tissue biopsies and post-mortem applications. Our data indicate that laboratory-based μCT as a modality can fill the gap between synchrotron radiation-based μCT and histology for a variety of tissues. As the information from haematoxylin and eosin (H&E) stained sections and μCT data is related, one can colourise local X-ray absorption values according to the H&E stain. Hence, μCT data can correlate and virtually extend two-dimensional (2D) histology data into the third dimension.
Biomaterials Science Center, Department of Biomedical Engineering, University of Basel, Allschwil, Switzerland.
Anna
A
Khimchenko
Biomaterials Science Center, Department of Biomedical Engineering, University of Basel, Allschwil, Switzerland.
Hans
H
Deyhle
Biomaterials Science Center, Department of Biomedical Engineering, University of Basel, Allschwil, Switzerland.
Georg
G
Schulz
Institute of Pathology, Department of Neuropathology, Basel University Hospital, Basel, Switzerland.
Gabriel
G
Schweighauser
Institute of Pathology, Department of Neuropathology, Basel University Hospital, Basel, Switzerland.
Jürgen
J
Hench
Biomaterials Science Center, Department of Biomedical Engineering, University of Basel, Allschwil, Switzerland; Medical Image Analysis Center, Department of Biomedical Engineering, University of Basel, Allschwil, Switzerland.
Natalia
N
Chicherova
Biomaterials Science Center, Department of Biomedical Engineering, University of Basel, Allschwil, Switzerland.
Christos
C
Bikis
Biomaterials Science Center, Department of Biomedical Engineering, University of Basel, Allschwil, Switzerland.
Simone E
SE
Hieber
Biomaterials Science Center, Department of Biomedical Engineering, University of Basel, Allschwil, Switzerland. Electronic address: bert.mueller@unibas.ch.
Bert
B
Müller
D000368:Aged; D000465:Algorithms; D001921:Brain / Q000000981:diagnostic imaging* / Q000473:pathology*; D002102:Cadaver; D016591:Histocytological Preparation Techniques / Q000379:methods*; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008297:Male; D011856:Radiographic Image Enhancement / Q000379:methods; D011857:Radiographic Image Interpretation, Computer-Assisted / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D055114:X-Ray Microtomography / Q000379:methods*
2D-3D image registration; Hard X-ray tomography; Histology; Joint histogram analysis; Nano-focus X-ray; Paraffin-embedded human cerebellum; Purkinje cells