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Three-dimensional visualization of cerebral blood vessels and neural changes in thick ischemic rat brain slices using tissue clearing

 Eun-Joo Lee  ;  Sung-Kuk Hong  ;  Dong-Hwa Choi  ;  Sang-Il Gum  ;  Mee Yul Hwang  ;  Dong Sun Kim  ;  Ji Won Oh  ;  Eun-Shil Lee 
 SCIENTIFIC REPORTS, Vol.12(1) : 15897, 2022-09 
Journal Title
Issue Date
Animals ; Brain* / pathology ; Imaging, Three-Dimensional* / methods ; Infarction, Middle Cerebral Artery / diagnostic imaging ; Infarction, Middle Cerebral Artery / pathology ; Ischemia / pathology ; Lectins ; Microscopy, Confocal ; Rats
Blood vessels are three-dimensional (3D) in structure and precisely connected. Conventional histological methods are unsuitable for their analysis because of the destruction of functionally important topological 3D vascular structures. Tissue optical clearing techniques enable extensive volume imaging and data analysis without destroying tissue. This study therefore applied a tissue clearing technique to acquire high-resolution 3D images of rat brain vasculature using light-sheet and confocal microscopies. Rats underwent middle cerebral artery occlusion for 45 min followed by 24 h reperfusion with lectin injected directly into the heart for vascular staining. For acquiring 3D images of rat brain vasculature, 3-mm-thick brain slices were reconstructed using tissue clearing and light-sheet microscopy. Subsequently, after 3D rendering, the fitting of blood vessels to a filament model was used for analysis. The results revealed a significant reduction in vessel diameter and density in the ischemic region compared to those in contralesional non-ischemic regions. Immunostaining of 0.5-mm-thick brain slices revealed considerable neuronal loss and increased astrocyte fluorescence intensity in the ipsilateral region. Thus, these methods can provide more accurate data by broadening the scope of the analyzed regions of interest for examining the 3D cerebrovascular system and neuronal changes occurring in various brain disorders.
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1. College of Medicine (의과대학) > Dept. of Anatomy (해부학교실) > 1. Journal Papers
Yonsei Authors
Oh, Ji Won(오지원) ORCID logo https://orcid.org/0000-0001-5742-5120
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