Xue Yi, Berry Kalen P, Boivin Josiah R, Wadduwage Dushan, Nedivi Elly, So Peter T C
Department of Mechanical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA 02139, USA.
Laser Biomedical Research Center, 77 Massachusetts Ave., Cambridge, MA 02139, USA.
Biomed Opt Express. 2018 Oct 22;9(11):5654-5666. doi: 10.1364/BOE.9.005654. eCollection 2018 Nov 1.
Line-scanning temporal focusing microscopy (LineTFM) is capable of imaging biological samples more than 10 times faster than two-photon laser point-scanning microscopy (TPLSM), while achieving nearly the same lateral and axial spatial resolution. However, the image contrast taken by LineTFM is lower than that by TPLSM because LineTFM is severely influenced by biological tissue scattering. To reject the scattered photons, we implemented LineTFM using both structured illumination and uniform illumination combined with the HiLo post-processing algorithm, called HiLL microscopy (HiLo-Line-scanning temporal focusing microscopy). HiLL microscopy significantly reduces tissue scattering and improves image contrast. We demonstrate HiLL microscopy with brain imaging. This approach could potentially find applications in monitoring fast dynamic events and in mapping high resolution structures over a large volume.
线扫描时间聚焦显微镜（LineTFM）能够以比双光子激光点扫描显微镜（TPLSM）快10倍以上的速度对生物样本进行成像，同时实现几乎相同的横向和轴向空间分辨率。然而，LineTFM拍摄的图像对比度低于TPLSM，因为LineTFM受到生物组织散射的严重影响。为了去除散射光子，我们采用结构化照明和均匀照明相结合，并结合HiLo后处理算法实现了LineTFM，即HiLL显微镜（HiLo线扫描时间聚焦显微镜）。HiLL显微镜显著减少了组织散射并提高了图像对比度。我们通过脑成像展示了HiLL显微镜。这种方法可能在监测快速动态事件以及在大体积范围内绘制高分辨率结构方面找到应用。
