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A combinatorial method to visualize the neuronal network in the mouse spinal cord: combination of a modified Golgi-Cox method and synchrotron radiation micro-computed tomography.

ABSTRACT: Exploring the three-dimensional (3D) morphology of neurons is essential to understanding spinal cord function and associated diseases comprehensively. However, 3D imaging of the neuronal network in the broad region of the spinal cord at cellular resolution remains a challenge in the field of neuroscience. In this study, to obtain high-resolution 3D imaging of a detailed neuronal network in the mass of the spinal cord, the combination of synchrotron radiation micro-computed tomography (SRμCT) and the Golgi-cox staining were used. We optimized the Golgi-Cox method (GCM) and developed a modified GCM (M-GCM), which improved background staining, reduced the number of artefacts, and diminished the impact of incomplete vasculature compared to the current GCM. Moreover, we achieved high-resolution 3D imaging of the detailed neuronal network in the spinal cord through the combination of SRμCT and M-GCM. Our results showed that the M-GCM increased the contrast between the neuronal structure and its surrounding extracellular matrix. Compared to the GCM, the M-GCM also diminished the impact of the artefacts and incomplete vasculature on the 3D image. Additionally, the 3D neuronal architecture was successfully quantified using a combination of SRμCT and M-GCM. The SRμCT was shown to be a valuable non-destructive tool for 3D visualization of the neuronal network in the broad 3D region of the spinal cord. Such a combinatorial method will, therefore, transform the presentation of Golgi staining from 2 to 3D, providing significant improvements in the 3D rendering of the neuronal network.

SUBMITTER: Jiang L

PROVIDER: S-EPMC8062354 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:This study used available or purchased equipment and an image reconstruction system developed by the college of dentistry to establish a basic self-assembled micro-computed tomography (micro-CT) system. Such a system would be suitable for teaching dental radiology to dental students. Specifically, it could help students to understand the principles governing dental cone-beam computed tomography (CBCT) and provide graduate students with a system for scanning small samples (e.g., individual teeth) during the early stages of research. The self-assembled micro-CT system was constructed using a portable dental X-ray tube, an intraoral digital X-ray detector, a high-precision rotation stage, related bracket accessories, and a notebook computer. Reconstructed images and three-dimensional models of the maxillary right third molar were produced using the self-assembled micro-CT system and an advanced commercially available micro-CT system (Skyscan 2211). Subsequently, the reconstructed images and 3D models produced using the two systems were compared by two senior dentists to determine whether considerable visual differences could be observed. Finally, the signal-to-noise ratio (SNR) was used for quantitative analysis and to compare the systems. Although the self-assembled micro-CT system produced image boundaries that were not as sharp as those of Skyscan 2211, the images were nonetheless remarkably similar. In addition, the two micro-CT systems produced 3D models that were almost identical in appearance and root canal shape. Quantitative analysis revealed that Skyscan 2211 had produced a SNR that was superior to that of the self-assembled micro-CT system, with the difference ranging from 36.77% to 136.22%; enamel, which has a higher density, exhibited lower SNR differences, whereas dentin, which has a lower density, exhibited higher SNR differences. The self-assembled micro-CT system with a resolution of 36 μm was created using a portable dental X-ray tube and an intraoral digital X-ray detector. Although the scanning time was relatively long (~30 min to scan images of a tooth), the images were adequate in the preliminary stage of experiments. More importantly, students were afforded the opportunity to observe the process of assembling and disassembling each component of a micro-CT scanner and thereby achieve a more comprehensive understanding of the principles governing micro-CT and dental CBCT.

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A combinatorial method to visualize the neuronal network in the mouse spinal cord: combination of a modified Golgi-Cox method and synchrotron radiation micro-computed tomography.

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