4.7 Article

Slide-free histological imaging by microscopy with ultraviolet surface excitation using speckle illumination

Journal

PHOTONICS RESEARCH
Volume 10, Issue 1, Pages 120-125

Publisher

CHINESE LASER PRESS
DOI: 10.1364/PRJ.440935

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Funding

  1. Research Grants Council of the Hong Kong Special Administrative Region [26203619, 16208620]
  2. Hong Kong Innovation and Technology Commission [ITS/036/19]

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Microscopy with ultraviolet surface excitation (MUSE) is a promising slide-free imaging technique that improves the histopathology workflow. This study proposes the incorporation of MUSE with speckle illumination (MUSES) to achieve sharp imaging on thick and rough specimens with improved contrast. Experimental results demonstrate the potential of MUSES in providing histological images with spatial resolution of around 1 µm within 10 minutes.
Microscopy with ultraviolet surface excitation (MUSE) is a promising slide-free imaging technique to improve the time-consuming histopathology workflow. However, since the penetration depth of the excitation light is tissue dependent, the image contrast could be significantly degraded when the depth of field of the imaging system is shallower than the penetration depth. High-resolution cellular imaging normally comes with a shallow depth of field, which also restricts the tolerance of surface roughness in biological specimens. Here we propose the incorporation of MUSE with speckle illumination (termed MUSES), which can achieve sharp imaging on thick and rough specimens. Our experimental results demonstrate the potential of MUSES in providing histological images with similar to 1 mu m spatial resolution and improved contrast, within 10 minutes for a field of view of 1.7 mm x 1.2 mm. With the extended depth of field feature, MUSES also relieves the constraint of tissue flatness. Furthermore, with a color transformation assisted by deep learning, a virtually stained histological image can be generated without manual tuning, improving the applicability of MUSES in clinical settings. (C) 2021 Chinese Laser Press

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