Ahn Soyeon, Yoon Jung-Yeon, Kim Pilhan
R&D Center, IVIM Technology, 17 Techno 4-ro, Yuseong-gu, Daejeon, 34013, Republic of Korea.
Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.
Eur Heart J Imaging Methods Pract. 2024 Jul 1;2(1):qyae062. doi: 10.1093/ehjimp/qyae062. eCollection 2024 Jan.
To develop and validate an optimized intravital heart microimaging protocol using a suction-based tissue motion-stabilizing cardiac imaging window to facilitate real-time observation of dynamic cellular behaviours within cardiac tissue in live mouse models.
Intravital heart imaging was conducted using dual-mode confocal and two-photon microscopy. Mice were anesthetized, intubated, and maintained at a stable body temperature during the procedure. LysM-eGFP transgenic mice were utilized to visualize immune cell dynamics with vascular labelling by intravenous injection of anti-CD31 antibody and DiD-labelled red blood cells (RBCs). A heart imaging window chamber with a vacuum-based tissue motion stabilizer with 890-920 mbar was applied following a chest incision to expose the cardiac tissue. The suction-based heart imaging window chamber system and artificial intelligence-based motion compensation function significantly reduced motion artefacts and facilitated real-time cell analysis of immune cell and RBC trafficking, revealing a mean neutrophil movement velocity of 1.66 mm/s, which was slower compared to the RBC flow velocity of 9.22 mm/s. Intravital two-photon microscopic heart imaging enabled label-free second harmonic generation imaging of cardiac muscle structures with 820-840 nm excitation wavelength, revealing detailed biodistributions and structural variations in sarcomeres and fibrillar organization in the heart.
The optimized intravital heart imaging protocol successfully demonstrates its capability to provide high-resolution, real-time visualization of dynamic cellular activities within live cardiac tissue.
开发并验证一种优化的活体心脏显微成像方案，该方案使用基于吸力的组织运动稳定心脏成像窗口，以促进在活体小鼠模型中实时观察心脏组织内动态细胞行为。
使用双模式共聚焦显微镜和双光子显微镜进行活体心脏成像。在操作过程中，小鼠被麻醉、插管并维持稳定体温。利用LysM-eGFP转基因小鼠，通过静脉注射抗CD31抗体和DiD标记的红细胞（RBC）进行血管标记，以可视化免疫细胞动态。胸部切开后，应用具有890 - 920毫巴真空组织运动稳定器的心脏成像窗口腔室来暴露心脏组织。基于吸力的心脏成像窗口腔室系统和基于人工智能的运动补偿功能显著减少了运动伪影，并有助于对免疫细胞和红细胞运输进行实时细胞分析，结果显示中性粒细胞平均移动速度为1.66毫米/秒，与红细胞流速9.22毫米/秒相比更慢。活体双光子显微镜心脏成像能够在820 - 840纳米激发波长下对心肌结构进行无标记二次谐波生成成像，揭示了心脏中肌节和纤维组织的详细生物分布和结构变化。
优化的活体心脏成像方案成功展示了其在活体心脏组织中提供高分辨率、实时动态细胞活动可视化的能力。
