title: Extending Whole Slide Imaging: Color Darkfield Internal Reflection Illumination (DIRI) for Biological Applications.
abstract: 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.
authors.affiliation: The Department of Biomedical Engineering, Graduate School of Biomedical Engineering, Tohoku University, Sendai, Miyagi, Japan.|Olympus Corporation, Shinjuku-Ku, Tokyo, Japan.
authors.forename: Yoshihiro
authors.identifier: 0000-0001-6442-8733
authors.initials: Y
authors.lastname: Kawano
authors.affiliation: Cell Function & Dynamics, Brain Science Institute, RIKEN, Wako, Saitama, Japan.
authors.forename: Kana
authors.initials: K
authors.lastname: Namiki
authors.affiliation: Cell Function & Dynamics, Brain Science Institute, RIKEN, Wako, Saitama, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Miyawaki
authors.affiliation: The Department of Biomedical Engineering, Graduate School of Biomedical Engineering, Tohoku University, Sendai, Miyagi, Japan.|Department of Finemechanics, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Takuji
authors.initials: T
authors.lastname: Ishikawa
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D001921:Brain / Q000166:cytology*; D001931:Brain Mapping / Q000379:methods*; D005056:Euglena gracilis / Q000166:cytology* / Q000378:metabolism; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008027:Light; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria