平场定量相位显微镜在骨髓间充质干细胞线粒体动力学观察中的应用研究(特邀)  

Application of Flat-Field Quantitative Phase Microscopy for Observing Mitochondrial Dynamics in Bone Marrow Mesenchymal Stem Cells (Invited)

作　　者：戴太强 马英 杜宇轩 侯燕 吕前欣 康娟 姚保利[6] 郜鹏 孔亮 Dai Taiqiang;Ma Ying;Du Yuxuan;Hou Yan;Li Qianxnin;Kang Juan;Yao Baoli;Gao Peng;Kong Liang(State Key Laboratory of Oral&Macillofacial Reconstruction and Regeneration,Xi'an 710032,Shaanci,China;National Clinical Research Center for Oral Diseases,Xi'an 710032,Shaanci,China;Shaani Clinical Research Center for Oral Diseases,Department of Oral,Xi'an 710032,Shaanxi,China;Mailofacial Surgery,School of Stomatology,Fourth Military Medical University,Xi'an 710032,Shaanci,China;School of Physics,Xidian University,Xi'an 710171,Shaanai,China;State Key Laboratory of Transient Optics and Photonics,Xi'an Institute of Optics and Precision Mechanics,Chinese Academy of Sciences,Xi'an 710119,Shaanxi,China)

机构地区：[1]口颌系统重建与再生全国重点实验室,陕西西安710032 [2]国家口腔疾病临床医学研究中心,陕西西安710032 [3]陕西省口腔疾病临床医学研究中心,陕西西安710032 [4]第四军医大学口腔医院颌面外科,陕西西安710032 [5]西安电子科技大学物理学院,陕西西安710171 [6]中国科学院西安光学精密机械研究所瞬态光学与光子技术国家重点实验室,陕西西安710119

出　　处：《激光与光电子学进展》2024年第6期308-314,共7页Laser & Optoelectronics Progress

基　　金：国家重点研发计划政府间国际科技创新合作(2022Y17E0l00700);空军军医大学伙伴实验室合作交流计划(2022HB011);陕西省自然科学基础研究计划项目(2022JQ-788)。

摘　　要：分离培养SD大鼠骨髓间充质干细胞,传代纯化后接种至共聚焦培养皿,置于自主研发搭建的平场定量相位显微镜下,利用荧光及相位双通道证实线粒体特征后,进行长时间无标记观察,分析平场定量相位显微镜观察到的线粒体分裂、融合过程,以及细胞凋亡过程中的线粒体变化。平场定量相位显微镜无标记观察到的线粒体可与荧光标记的线粒体完全重叠,表明平场定量相位显微镜可以清晰观察线粒体并进行无标记高分辨率成像。同时,平场定量相位显微镜可以对培养条件下的骨髓间充质干细胞进行长时间无标记观察,并高分辨率记录线粒体分裂、融合过程。此外,还利用平场定量相位显微镜首次完整记录了CCCP作用下线粒体的变化,该变化直观地呈现了线粒体途径的细胞凋亡过程。平场定量相位显微镜可以对培养的细胞进行长时间无标记高分辨率观察,为线粒体动力学的研究提供了一种新的观察手段。To investigate the feasibility of label-free imaging using flat-field quantitative phase microscopy for observing mitochondrial dynamics in mesenchymal stem cells(MSCs)derived from bone marrow,SD rat bone marrow MSCs were isolated and cultured.Following passages and purification,they were seeded into confocal culture dishes and placed under a flat-field quantitative phase microscope developed by our team.Label-free observations were conducted over an extended period after confirming mitochondrial characteristics using fluorescence and phase dual channels.The mitochondrial division,fusion processes,and mitochondrial changes during cell apoptosis observed using the flat-field quantitative phase microscope were analyzed.The mitochondria observed via unlabeled flat-field quantitative phase microscopy exhibited complete overlap with fluorescently labeled mitochondria,demonstrating the microscope's capability to visualize mitochondria and conduct unlabeled high-resolution imaging.Moreover,the flat-field quantitative phase microscope facilitated long-term unlabeled observations of bone marrow MSCs under cultivation conditions,enabling high-resolution recording of mitochondrial division and fusion processes.Furthermore,we used the flat-field quantitative phase microscope to record mitochondria changes under CCCP treatment for the first time,visually presenting the apoptosis via mitochondrial pathway.The flat-field quantitative phase microscope allows for prolonged,unlabeled,high-resolution observations of cultured cells,offering a new tool for investigating mitochondrial dynamics.

关 键 词：无标记成像 平场定量相位显微镜 骨髓间充质干细胞 线粒体动力学 

分 类 号：Q271[生物学—细胞生物学]