Chromatic confocal microscopy for multi-depth imaging of epithelial tissue.
Chromatic confocal microscopy for multi-depth imaging of epithelial tissue.
复制标题
DOI：
10.1364/boe.4.000732
复制
发表时间：
2013-05-01
影响因子：
3.4
通讯作者：
Maitland KC
中科院分区：
文献类型：
--
作者：
Olsovsky C;Shelton R;Carrasco-Zevallos O;Applegate BE;Maitland KC
我们提出了一种新的彩色共焦显微镜能够体积反射成像的微观结构在不透明的组织。我们的设计利用了非球面透镜的色差，否则很好地纠正。由多个非球面产生的强色差将超连续谱光纵向分散到样品上。因此，用光谱仪检测到的反向散射光被波长编码，并且每个光谱对应于线图像。这种方法消除了对传统轴向机械扫描技术的需要，传统轴向机械扫描技术难以实现内窥镜检查并且容易受到运动伪影的影响。波长范围为590-775 nm，成像深度>150 µm，轴向分辨率约为3 µm。该系统通过捕获颊粘膜的体积图像进一步证明。我们相信，这些代表了第一个显微结构图像在不透明的生物组织中使用彩色共聚焦显微镜，表现出长的成像深度，同时保持可接受的分辨率，解决细胞形态。该光学系统的小型化可以提高上皮组织的显微内窥镜体内体积成像的速度和精度。
We present a novel chromatic confocal microscope capable of volumetric reflectance imaging of microstructure in non-transparent tissue. Our design takes advantage of the chromatic aberration of aspheric lenses that are otherwise well corrected. Strong chromatic aberration, generated by multiple aspheres, longitudinally disperses supercontinuum light onto the sample. The backscattered light detected with a spectrometer is therefore wavelength encoded and each spectrum corresponds to a line image. This approach obviates the need for traditional axial mechanical scanning techniques that are difficult to implement for endoscopy and susceptible to motion artifact. A wavelength range of 590-775 nm yielded a >150 µm imaging depth with ~3 µm axial resolution. The system was further demonstrated by capturing volumetric images of buccal mucosa. We believe these represent the first microstructural images in non-transparent biological tissue using chromatic confocal microscopy that exhibit long imaging depth while maintaining acceptable resolution for resolving cell morphology. Miniaturization of this optical system could bring enhanced speed and accuracy to endomicroscopic in vivo volumetric imaging of epithelial tissue.