Multimodal label-free ex vivo imaging using a dual-wavelength microscope with axial chromatic aberration compensation
Multimodal label-free ex vivo imaging using a dual-wavelength microscope with axial chromatic aberration compensation
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DOI：
10.1117/1.jbo.23.9.091403
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发表时间：
2018-09-01
影响因子：
3.5
通讯作者：
Borile, Giulia
中科院分区：
文献类型：
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作者：
Filippi, Andrea;Dal Sasso, Eleonora;Borile, Giulia
无标记显微镜是一种非常强大的技术，可以应用于研究样品，而不需要外源荧光探针，保持多光子显微镜的主要优点，如更长的穿透深度和内在的光学切片，同时使同一标本上的系列多技术检查。在许多无标记的显微镜方法中，谐波发生（HG）是最有趣的方法之一，由于其通常具有较低的光毒性和相对容易实现。今天，HG和普通双光子显微镜（TPM）是成熟的技术，并经常用于几个研究领域。然而，它们需要大量的微调才能充分利用，这使得它们很难并行执行。在这里，我们介绍了我们设计的多模态显微镜，能够同时执行TPM和HG，而不需要任何妥协，这要归功于两个独立的，单独优化的激光源，具有轴向色差补偿。我们还将我们的设置应用于检查过多的离体样品，以证明其能力和多模式方法的显著优势。(C)2018年，摄影光学仪器工程师协会（SPIE）
Label-free microscopy is a very powerful technique that can be applied to study samples with no need for exogenous fluorescent probes, keeping the main benefits of multiphoton microscopy, such as longer penetration depths and intrinsic optical sectioning while enabling serial multitechniques examinations on the same specimen. Among the many label-free microscopy methods, harmonic generation (HG) is one of the most intriguing methods due to its generally low photo-toxicity and relative ease of implementation. Today, HG and common two-photon microscopy (TPM) are well-established techniques, and are routinely used in several research fields. However, they require a significant amount of fine-tuning to be fully exploited, making them quite difficult to perform in parallel. Here, we present our designed multimodal microscope, capable of performing simultaneously TPM and HG without any kind of compromise thanks to two, separate, individually optimized laser sources with axial chromatic aberration compensation. We also apply our setup to the examination of a plethora of ex vivo samples to prove its capabilities and the significant advantages of a multimodal approach. (C) 2018 Society of Photo-Optical Instrumentation Engineers (SPIE)