3D imaging of cleared ex vivo normal human skin, skin appendages and psoriasiform skin lesion using light-sheet microscopy (LSFM)
USING LIGHT-SHEET FLUORESCENCE MICROSCOPY (LSFM)
Research by C. Jardet (Genoskin), S. Abadie (Syntivia), M. Pastore (Genoskin), J. Colombelli (Advanced Digital Microscopy – IRB), B. Chaput (CHU Toulouse Rangueil), A. David (Genoskin), J.-L. Grolleau (CHU Toulouse Rangueil), P. Bedos (Syntivia), V. Lobjois (ITAV), P. Descargues (Genoskin), J. Roquette (ITAV)
Confocal and multiphoton microscopies have emerged as useful imaging tools for the non-invasive visualization of normal and pathological skin with high resolution. However, these techniques are characterized by a limited penetration depth due to light scattering and can cause photobleaching. Light-sheet fluorescence microscopy (LSFM) is an attractive approach that has shown promise for the acquisition of volumetric data of thick tissues. In this study, LSFM was combined with optical clearing methods to allow in-depth optical sectioning and generation of 3D images of entire human skin biopsies.
Skin topical clearing and skin imaging metholody
1. OPTICAL CLEARING OF HUMAN SKIN BIOPSIES
2. LSFM OF HUMAN SKIN BIOPSIES AND IMAGE PROCESSING
Volume rendering of a human skin with Amira® Software
Identification of morphological features associated with psoriasis in 3D
H&E staining of (A) healthy skin and (B) pathological skin. Epidermis segmentation and volume rendering of (C) healthy skin and (D) pathological skin following LSFM (excitation laser of 488 nm, emission filter of 525/50 nm). The volume of pathological skin epidermis (0,092 mm3) was almost twice as large as that of healthy skin (0,053 mm3). The pathological skin had a thickness of 203,7 μm compared to 117,9 μm for healthy skin. Psoriasiform skin lesion was obtained from a female donor. Scale bar: 100 μm.