Wang Zhi, Xie Ruiheng, Shi Qishuo, Li Yafeng, Chang Jin, Yuan Jing, Gong Hui, Chen Jianwei
Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, MoE Key Laboratory for Biomedical Photonics, Huazhong University of Science and Technology, Wuhan, China.
HUST-Suzhou Institute for Brainsmatics, JITRI, Suzhou, China.
Biomed Opt Express. 2023 May 4;14(6):2539-2550. doi: 10.1364/BOE.488766. eCollection 2023 Jun 1.
The use of combined optical imaging and tissue sectioning has potential for use in visualizing heart-wide fine structures at single-cell resolution. However, existing tissue preparation methods fail to generate ultrathin cavity-containing cardiac tissue slices with minimal deformation. This study developed an efficient vacuum-assisted tissue embedding method to prepare high-filled, agarose-embedded whole-heart tissue. Utilizing optimized vacuum parameters, we achieved 94% filled whole-heart tissue with the thinnest cut slice of 5 µm. We subsequently imaged a whole mouse heart sample using vibratome-integrated fluorescence micro-optical sectioning tomography (fMOST) with a voxel size of 0.32 µm × 0.32 µm × 1 µm. The imaging results indicated that the vacuum-assisted embedding method enabled whole-heart tissue to withstand long-term thin cutting while ensuring that slices were consistent and of high quality.
联合光学成像和组织切片技术在以单细胞分辨率可视化全心脏精细结构方面具有应用潜力。然而，现有的组织制备方法无法生成变形最小的超薄含腔心脏组织切片。本研究开发了一种高效的真空辅助组织包埋方法，以制备高填充度、琼脂糖包埋的全心脏组织。利用优化的真空参数，我们获得了填充度为94%的全心脏组织，最薄切片厚度为5 µm。随后，我们使用体素大小为0.32 µm × 0.32 µm × 1 µm的振动切片机集成荧光显微光学切片断层扫描（fMOST）对整个小鼠心脏样本进行成像。成像结果表明，真空辅助包埋方法使全心脏组织能够承受长期的薄切片，同时确保切片一致且质量高。
