TY - JOUR
T1 - Extending whole slide imaging
T2 - Color darkfield internal reflection illumination (DIRI) for biological applications
AU - Kawano, Yoshihiro
AU - Namiki, Kana
AU - Miyawaki, Atsushi
AU - Ishikawa, Takuji
N1 - Funding Information:
This study was supported by a Grant-in-Aid for Scientific Research (A), No. 26242039 ( http://www.jsps.go.jp/english/e-grants/index.html ). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. We would like to thank Brendan Brinkman of Olympus Corporation for his advice on this article. Olympus Corporation provided several microscope parts, but did not fund this study nor influence its outcome.
Publisher Copyright:
© 2017 Kawano et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2017/1/1
Y1 - 2017/1/1
N2 - 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.
AB - 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.
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U2 - 10.1371/journal.pone.0167774
DO - 10.1371/journal.pone.0167774
M3 - Article
C2 - 28085892
AN - SCOPUS:85009250393
SN - 1932-6203
VL - 12
JO - PLoS One
JF - PLoS One
IS - 1
M1 - e0167774
ER -