Britton Chance Center and MoE Key Laboratory for Biomedical Photonics, Advanced Biomedical Imaging Facility, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan 430074, Hubei, China.
Changping Laboratory, Beijing 102206, China.
J Phys Chem B. 2023 May 18;127(19):4229-4234. doi: 10.1021/acs.jpcb.2c09114. Epub 2023 May 4.
High-throughput stimulated Raman scattering (SRS) microscopy is highly desired for large tissue imaging with chemical specificity. However, the mapping speed remains as the major short board of conventional SRS, primarily owing to the mechanical inertia existing in galvanometers or other laser scanning alternatives. Here, we developed inertia-free acousto-optic deflector (AOD)-based high-speed large-field stimulated Raman scattering microscopy, in which both the speed and integration time are ensured by immune of the mechanical response time. To avoid laser beam distortion induced by the intrinsic spatial dispersion of AODs, two spectral compression systems are implemented to compress the broad-band femtosecond pulse to picosecond laser. We achieved an SRS imaging of a 12 × 8 mm mouse brain slice in only 8 min at an image resolution of approximately 1 μm and 32 slices from a whole brain in 12 h. The AOD-based inertia-free SRS mapping can be much faster after further upgrading and allow broad-spectrum applications of chemical imaging in the future.
高通量受激拉曼散射（SRS）显微镜在具有化学特异性的大组织成像方面有很高的需求。然而，由于传统 SRS 中存在的机械惯性，主要是由于检流计或其他激光扫描替代方案中的机械惯性，其映射速度仍然是主要的短板。在这里，我们开发了基于无惯性声光偏转器（AOD）的高速大视场受激拉曼散射显微镜，其中通过免疫机械响应时间来确保速度和积分时间。为了避免 AOD 固有空间色散引起的激光束变形，实现了两个光谱压缩系统，将宽带飞秒脉冲压缩到皮秒激光。我们仅用 8 分钟就实现了 12×8mm 大小的老鼠脑切片的 SRS 成像，图像分辨率约为 1μm，用 12 小时从整个大脑获得 32 个切片。在进一步升级后，基于 AOD 的无惯性 SRS 映射速度可以更快，并允许在未来进行更广泛的化学成像应用。
