Three-Dimensional Imaging of Pulmonary Fibrotic Foci at the Alveolar Scale Using Tissue-Clearing Treatment with Staining Techniques of Extracellular Matrix.

IF 3.3 Q2 ENGINEERING, BIOMEDICAL International Journal of Biomedical Imaging Pub Date : 2020-12-29 eCollection Date: 2020-01-01 DOI:10.1155/2020/8815231
Kohei Togami, Hiroaki Ozaki, Yuki Yumita, Anri Kitayama, Hitoshi Tada, Sumio Chono
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引用次数: 3

Abstract

Idiopathic pulmonary fibrosis is a progressive, chronic lung disease characterized by the accumulation of extracellular matrix proteins, including collagen and elastin. Imaging of extracellular matrix in fibrotic lungs is important for evaluating its pathological condition as well as the distribution of drugs to pulmonary focus sites and their therapeutic effects. In this study, we compared techniques of staining the extracellular matrix with optical tissue-clearing treatment for developing three-dimensional imaging methods for focus sites in pulmonary fibrosis. Mouse models of pulmonary fibrosis were prepared via the intrapulmonary administration of bleomycin. Fluorescent-labeled tomato lectin, collagen I antibody, and Col-F, which is a fluorescent probe for collagen and elastin, were used to compare the imaging of fibrotic foci in intact fibrotic lungs. These lung samples were cleared using the ClearT2 tissue-clearing technique. The cleared lungs were two dimensionally observed using laser-scanning confocal microscopy, and the images were compared with those of the lung tissue sections. Moreover, three-dimensional images were reconstructed from serial two-dimensional images. Fluorescent-labeled tomato lectin did not enable the visualization of fibrotic foci in cleared fibrotic lungs. Although collagen I in fibrotic lungs could be visualized via immunofluorescence staining, collagen I was clearly visible only until 40 μm from the lung surface. Col-F staining facilitated the visualization of collagen and elastin to a depth of 120 μm in cleared lung tissues. Furthermore, we visualized the three-dimensional extracellular matrix in cleared fibrotic lungs using Col-F, and the images provided better visualization than immunofluorescence staining. These results suggest that ClearT2 tissue-clearing treatment combined with Col-F staining represents a simple and rapid technique for imaging fibrotic foci in intact fibrotic lungs. This study provides important information for imaging various organs with extracellular matrix-related diseases.

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利用细胞外基质染色技术组织清除处理肺泡尺度肺纤维化病灶的三维成像。
特发性肺纤维化是一种进行性慢性肺部疾病,其特征是细胞外基质蛋白(包括胶原蛋白和弹性蛋白)的积累。纤维化肺的细胞外基质成像对于评价其病理状况、药物在肺病灶部位的分布及其治疗效果具有重要意义。在这项研究中,我们比较了细胞外基质染色技术和光学组织清除治疗技术,以开发肺纤维化病灶部位的三维成像方法。通过肺内给药博来霉素制备肺纤维化小鼠模型。采用荧光标记的番茄凝集素、I型胶原抗体和Col-F(胶原蛋白和弹性蛋白的荧光探针)比较完整纤维化肺中纤维化灶的影像学表现。使用ClearT2组织清除技术清除这些肺样本。用激光共聚焦显微镜对清除后的肺进行二维观察,并与肺组织切片图像进行比较。此外,将连续二维图像重构为三维图像。荧光标记的番茄凝集素不能在清除的纤维化肺中显示纤维化灶。虽然通过免疫荧光染色可以看到纤维化肺中的胶原I,但直到距离肺表面40 μm时才清晰可见胶原I。在清除后的肺组织中,Col-F染色使胶原蛋白和弹性蛋白的可见深度达到120 μm。此外,我们使用Col-F可视化清除纤维化肺的三维细胞外基质,其图像比免疫荧光染色提供更好的可视化效果。这些结果表明,在完整的纤维化肺中,ClearT2组织清除治疗联合Col-F染色是一种简单快速的成像纤维化灶的技术。本研究为细胞外基质相关疾病的各种器官成像提供了重要信息。
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来源期刊
CiteScore
12.00
自引率
0.00%
发文量
11
审稿时长
20 weeks
期刊介绍: The International Journal of Biomedical Imaging is managed by a board of editors comprising internationally renowned active researchers. The journal is freely accessible online and also offered for purchase in print format. It employs a web-based review system to ensure swift turnaround times while maintaining high standards. In addition to regular issues, special issues are organized by guest editors. The subject areas covered include (but are not limited to): Digital radiography and tomosynthesis X-ray computed tomography (CT) Magnetic resonance imaging (MRI) Single photon emission computed tomography (SPECT) Positron emission tomography (PET) Ultrasound imaging Diffuse optical tomography, coherence, fluorescence, bioluminescence tomography, impedance tomography Neutron imaging for biomedical applications Magnetic and optical spectroscopy, and optical biopsy Optical, electron, scanning tunneling/atomic force microscopy Small animal imaging Functional, cellular, and molecular imaging Imaging assays for screening and molecular analysis Microarray image analysis and bioinformatics Emerging biomedical imaging techniques Imaging modality fusion Biomedical imaging instrumentation Biomedical image processing, pattern recognition, and analysis Biomedical image visualization, compression, transmission, and storage Imaging and modeling related to systems biology and systems biomedicine Applied mathematics, applied physics, and chemistry related to biomedical imaging Grid-enabling technology for biomedical imaging and informatics
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