You Sixian, Sun Yi, Chaney Eric J, Zhao Youbo, Chen Jianxin, Boppart Stephen A, Tu Haohua
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Biomed Opt Express. 2018 Oct 5;9(11):5240-5252. doi: 10.1364/BOE.9.005240. eCollection 2018 Nov 1.
Histochemistry is a microscopy-based technology widely used to visualize the molecular distribution in biological tissue. Recent developments in label-free optical imaging has demonstrated the potential to replace the conventional histochemical labels/markers (fluorescent antibodies, organic dyes, nucleic acid probes, and other contrast agents) with diverse optical interactions to generate histochemical contrasts, allowing "virtual" histochemistry in three spatial dimensions without preparing a microscope slide (i.e. labor-intensive sample preparation). However, the histochemical information in a label-free optical image has often been rather limited due to the difficulty in simultaneously generating multiple histochemical contrasts with strict spatial co-registration. Here, in the first part (Part I) of this two-part series study, we develop a technique of slide-free virtual histochemistry based on label-free multimodal multiphoton microscopy, and simultaneously generate up to four histochemical contrasts from animal and human tissue. To enable this functionality, we construct and demonstrate a robust fiber-based laser source for clinical translation and phenotype a wide variety of vital cells in unperturbed mammary tissue.
组织化学是一种基于显微镜的技术，广泛用于可视化生物组织中的分子分布。无标记光学成像的最新进展已证明，利用多种光学相互作用来产生组织化学对比度，从而取代传统的组织化学标记/标志物（荧光抗体、有机染料、核酸探针和其他造影剂）的潜力，这使得无需制备显微镜载玻片（即劳动密集型的样品制备）就能在三个空间维度上实现“虚拟”组织化学。然而，由于难以同时产生具有严格空间共配准的多种组织化学对比度，无标记光学图像中的组织化学信息往往相当有限。在此，在这个两部分系列研究的第一部分（第一部分）中，我们开发了一种基于无标记多模态多光子显微镜的无载玻片虚拟组织化学技术，并同时从动物和人体组织中产生多达四种组织化学对比度。为实现这一功能，我们构建并展示了一种用于临床转化的强大的基于光纤的激光源，并对未受干扰的乳腺组织中的多种活细胞进行表型分析。
