Research Techniques Made Simple: Optical Clearing and Three-Dimensional Volumetric Imaging of Skin Biopsies

June 2020 in “ Journal of Investigative Dermatology ”

Yingrou Tan, Carolyn Chiam, Yuning Zhang, Hong Liang Tey, Lai Guan Ng

three-dimensional volumetric imaging skin histology hair follicles sebaceous glands optical clearing solvent-based approaches aqueous-based approaches fixation permeabilization immunostaining light-sheet fluorescence microscope two-photon microscope melanin 3D imaging skin biopsy oil glands solvent-based methods water-based methods light-sheet microscope

TLDR 3D imaging of skin biopsies offers better accuracy but is time-consuming and can't clear melanin.

The document discussed the limitations of traditional two-dimensional skin histology and the advantages of three-dimensional (3D) volumetric imaging for accurately representing skin structures such as nerves, vasculature, hair follicles, and sebaceous glands. It highlighted the technical challenges of imaging deep into tissue due to light scattering and refractive index mismatches. Recent developments in optical clearing methods, which render tissues transparent, were reviewed, including solvent-based and aqueous-based approaches. The tissue preparation workflow, involving steps like fixation, permeabilization, and immunostaining, was outlined as crucial for high-quality imaging. The choice of microscope, such as light-sheet fluorescence or two-photon microscopes, depends on the required resolution. Despite the benefits of 3D imaging, limitations include the lengthy and laborious tissue preparation process and the inability to clear melanin in pigmented skin.

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