利用波前调制技术提升光透明样品的双光子成像分辨率

发布时间：2018-11-04 15:25

【摘要】：组织光透明技术结合双光子显微成像(TPM)技术能够有效提升生物样品的显微成像深度,然而现有的光透明剂与常用显微物镜的浸润介质折射率并不匹配,会引入球差从而降低深层组织成像的荧光强度和分辨率。针对该问题分析了球差对TPM荧光强度和分辨率的影响,建立了由物镜特性(数值孔径和浸润介质)、聚焦深度和物体折射率等参数构成的球差补偿模型,进而指导空间光调制器进行球差补偿。对荧光小球仿体样品和光透明脑组织样品的双光子成像结果显示,该球差补偿方法能显著提升样品信号量和系统纵向分辨率。另外,该方法在校正过程中无需多次成像,操作简单且耗时短,对光透明剂和显微物镜无特殊要求,具有较强的通用性。
[Abstract]:Tissue transparency technology combined with two-photon microscopic imaging (TPM) technique can effectively enhance the depth of microscopic imaging of biological samples. However, the refractive index of the existing optical transparency agent does not match the infiltrating medium index of the common objective lens. Spherical aberration is introduced to reduce the fluorescence intensity and resolution of deep tissue imaging. The effect of spherical aberration on the fluorescence intensity and resolution of TPM is analyzed, and a spherical compensation model is established, which is composed of objective properties (numerical aperture and infiltrating medium), focusing depth and refractive index of object. Then guide the spatial light modulator to compensate the spherical aberration. The results of two-photon imaging of fluorescent microspheres and light transparent brain tissue samples show that the spherical aberration compensation method can significantly improve the signal quantity of the sample and the longitudinal resolution of the system. In addition, the method does not need multiple imaging in the correction process, and the operation is simple and time consuming is short. The method has no special requirements for optical transparency agent and micro objective lens, so it has strong generality.
【作者单位】：哈尔滨工业大学深圳研究生院电子与信息工程学院;中国科学院深圳先进技术研究院生物医学光学与分子影像研究室;
【基金】：国家自然科学基金(81471702) 深圳市科技创新委员会项目(KQCX20120816155352228,JCYJ20140610153345808)
【分类号】：O439

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