用统一的方法校正色散和像差的显微OCT的相干性和衍射极限分辨率

H. Schulz-Hildebrandt, M. Münter, M. Ahrens, H. Spahr, Dierck Hillmann, P. König, G. Hüttmann
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引用次数: 4

摘要

光学相干断层扫描(OCT)图像散射组织5至15 μm分辨率。这通常不足以区分细胞和亚细胞结构。提高轴向和横向分辨率,并补偿由色散和像差引起的伪影,需要实现细胞和亚细胞分辨率。这包括在高横向分辨率下限制可用景深的离焦。OCT可以获得散射光的相位,因此可以通过数值算法来校正色散和像差。本文提出了一种基于相位误差多项式参数化和香农熵图像质量优化的色散/像差统一校正方法。为了验证,我们将超连续光源和400 nm带宽的自制光谱仪与高NA显微镜物镜结合在一起,用于组织和小动物成像。利用这种设置和计算校正,可以实现1.5 μm分辨率的体积成像。细胞和近细胞分辨率是证明在猪角膜和果蝇幼虫,当计算校正色散和像差是使用。由于使用的显微镜物镜的良好的校正,离焦是造成像差的主要原因。此外,还成功地校正了样品本身引起的高像差。在显微OCT成像中,色散和像差是密切相关的伪影。因此,它们可以通过优化图像质量以同样的方式进行校正。这种显微分辨率很容易在静态生物组织的OCT成像中实现。
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Coherence and diffraction limited resolution in microscopic OCT by a unified approach for the correction of dispersion and aberrations
Optical coherence tomography (OCT) images scattering tissues with 5 to 15 μm resolution. This is usually not sufficient for a distinction of cellular and subcellular structures. Increasing axial and lateral resolution and compensation of artifacts caused by dispersion and aberrations is required to achieve cellular and subcellular resolution. This includes defocus which limit the usable depth of field at high lateral resolution. OCT gives access the phase of the scattered light and hence correction of dispersion and aberrations is possible by numerical algorithms. Here we present a unified dispersion/aberration correction which is based on a polynomial parameterization of the phase error and an optimization of the image quality using Shannon’s entropy. For validation, a supercontinuum light sources and a costume-made spectrometer with 400 nm bandwidth were combined with a high NA microscope objective in a setup for tissue and small animal imaging. Using this setup and computation corrections, volumetric imaging at 1.5 μm resolution is possible. Cellular and near cellular resolution is demonstrated in porcine cornea and the drosophila larva, when computational correction of dispersion and aberrations is used. Due to the excellent correction of the used microscope objective, defocus was the main contribution to the aberrations. In addition, higher aberrations caused by the sample itself were successfully corrected. Dispersion and aberrations are closely related artifacts in microscopic OCT imaging. Hence they can be corrected in the same way by optimization of the image quality. This way microscopic resolution is easily achieved in OCT imaging of static biological tissues.
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