Kawano Yoshihiro, Namiki Kana, Miyawaki Atsushi, Ishikawa Takuji
The Department of Biomedical Engineering, Graduate School of Biomedical Engineering, Tohoku University, Sendai, Miyagi, Japan.
Olympus Corporation, Shinjuku-Ku, Tokyo, Japan.
PLoS One. 2017 Jan 13;12(1):e0167774. doi: 10.1371/journal.pone.0167774. eCollection 2017.
Whole slide imaging (WSI) is a useful tool for multi-modal imaging, and in our work, we have often combined WSI with darkfield microscopy. However, traditional darkfield microscopy cannot use a single condenser to support high- and low-numerical-aperture objectives, which limits the modality of WSI. To overcome this limitation, we previously developed a darkfield internal reflection illumination (DIRI) microscope using white light-emitting diodes (LEDs). Although the developed DIRI is useful for biological applications, substantial problems remain to be resolved. In this study, we propose a novel illumination technique called color DIRI. The use of three-color LEDs dramatically improves the capability of the system, such that color DIRI (1) enables optimization of the illumination color; (2) can be combined with an oil objective lens; (3) can produce fluorescence excitation illumination; (4) can adjust the wavelength of light to avoid cell damage or reactions; and (5) can be used as a photostimulator. These results clearly illustrate that the proposed color DIRI can significantly extend WSI modalities for biological applications.
全玻片成像（WSI）是多模态成像的一种有用工具，在我们的工作中，我们经常将WSI与暗场显微镜相结合。然而，传统的暗场显微镜不能使用单个聚光镜来支持高数值孔径和低数值孔径物镜，这限制了WSI的模态。为了克服这一限制，我们之前开发了一种使用发光二极管（LED）的暗场内反射照明（DIRI）显微镜。尽管已开发的DIRI在生物学应用中很有用，但仍有一些重大问题有待解决。在本研究中，我们提出了一种名为彩色DIRI的新型照明技术。使用三色LED极大地提高了系统的能力，使得彩色DIRI能够：（1）优化照明颜色；（2）可与油浸物镜组合；（3）能产生荧光激发照明；（4）可调节光的波长以避免细胞损伤或反应；（5）可用作光刺激器。这些结果清楚地表明，所提出的彩色DIRI可以显著扩展WSI在生物学应用中的模态。
