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11TISSUE OPTICAL CLEARING: FROM IN VITRO TO IN VIVO
DAN ZHU1,2
1Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science
and Technology, China
2MoE Key Laboratory for Biomedical Photonics, Huazhong University of Science and Technology, China
Abstract
Biomedical photonics is currently one of the fastest growingfields of life sciences, which allows structural and functional analysis of tissues with high resolution and contrast unattainable by any other method [1-3]. However, the high scattering of turbid biological tissues limits the penetration of light, leading to strongly decreased imaging resolution and contrast as light propagates deeper into the tissue [4, 5]. Fortunately, novel tissue optical clearing technique could reduce the scattering of tissue and make it transparent for higher optical imaging quality [6-9]. This presentation will introduce the recently developed in vitro optical clearing methods for whole organs imaging, including FDISCO and MACS [10, 11]. And then I will demonstrate in vivo skull/skin optical clearing window for imaging structural and functional of cutaneous / cortical vascular and cells, also manipulating cortical vasculature [12-14].
Figure 1:LSFM imaging of neural structures in the mouse brain and gastrocnemius muscle after FDISCO clearing. (A) Image of the whole brain (Thy1 -GFP-M)cleared by FDISCO. (B) Comparison of the high-magnification images of the cleared brains assessed immediately after FDISCO, 3DISCO, and uDISCO clearing. The whitearrowheads mark the tiny nerve fibers detected. For different clearing methods, the same imaging parameters and image processing methods were used for the sameregions. (C) Images of cortical neurons in the FDISCO-cleared brain taken at 0 and 150 days after clearing, respectively. The neurons (e.g., white arrowheads) could stillbe viewed well after 150 days. (D) Fluorescence level quantification of cleared brains over time after FDISCO, 3DISCO, and uDISCO clearing (n = 4, 3, and 3, respectively). (E) 3D reconstruction and segmentation of nerve branches (green) and motor endplates (red) of the gastrocnemius muscle (Thy1-YFP-16) cleared by FDISCO. (F) Highmagnification images of the dashed boxed region in (E). [10]
Figure 2: MACS for imaging various organs and embryo of neural structures and function [11].
(a)
(b) (c)
Figure 3: (a) In vivo skin optical clearing for blood flow imaging[15]; (b) in vivo skull optical clearing for cortical neural imaging
and (c) cortical vascular functional imaging [12, 13, 15].
References
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5] X. F. Fan, W. T. Zheng, and D. J. Singh, Light scattering and surface plasmons on small spherical particles, Light-Science & Applications,3, e179, 2014.
6] T. Yu et al., Optical clearing for multiscale biological tissues, Journal of Biophotonics,11(2), e201700187, 2018.
7] R. Cai et al., Panoptic imaging of transparent mice reveals whole-body neuronal projections and skull-meninges connections, Nature Neuroscience,22(2), 317-327, 2019.
8] M. Belle et al., Tridimensional Visualization and Analysis of Early Human Development, Cell,169(1), 161-173 e112, 2017.
9] H. M. Lai et al., Next generation histology methods for three-dimensional imaging of fresh and archival human brain tissues, Nature Communications,9(1), 2018.
10] Y. Qi et al., FDISCO: Advanced solvent-based clearing method for imaging whole organs, Science advances,5(1), eaau8355-eaau8355, 2019.
11] J. T. Zhu et al., MACS: Rapid Aqueous Clearing System for 3D Mapping of Intact Organs, Advanced Science,7(8), 2020.
12] Y. J. Zhao et al., Skull optical clearing window for in vivo imaging of the mouse cortex at synaptic resolution, Light-Science & Applications,7(2), 17153, 2018.
13] C. Zhang et al., Age differences in photodynamic therapy-mediated opening of the blood-brain barrier through the optical clearing skull window in mice, Lasers in Surgery and Medicine,51(7), 625-633, 2019.
14] W. Feng et al., Comparison of cerebral and cutaneous microvascular dysfunction with the development of type 1 diabetes Theranostics,9(20), 5854-5868, 2019.
15] D. Zhu et al., Imaging dermal blood flow through the intact rat skin with an optical clearing method, J Biomed Opt,15(2), 026008, 2010.
