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.
Highlights
The complexity of the human body makes it difficult to understand
The color balance could be set on the color darkfield internal reflection illumination (DIRI) using a NeoPixel intensity interface where each color light-emitting diodes (LEDs) can be adjusted on a scale of intensity from 0 to 255, 255 being the maximum
We confirmed that DIRI with red, green, or blue LEDs properly illuminated the tissue microarray (TMA) sample with each color, and the DIRI image provided a more detailed structure than that with brightfield illumination with white light
Summary
The complexity of the human body makes it difficult to understand. It consists of between 50 and 75 trillion cells, and its genes are capable of creating over 20,000 proteins. Optical microscopy is the main tool used to investigate the structure and functions of cells, tissues, and organs in the human body. Scientists have a wide variety of imaging techniques and tools at their disposal to better understand complicated biological environments. Whole slide imaging (WSI) or so call vertual slide technology is one of the core imaging techniques, which enables automated imaging and allows scientists to observe large samples. The observation area of WSI is not limited to the field of view of an objective lens [1,2,3,4,5]
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