Deep-skin multiphoton microscopy in vivo excited at 1600 nm: A comparative investigation with silicone oil and deuterium dioxide immersion
Deep-skin multiphoton microscopy in vivo excited at 1600 nm: A comparative investigation with silicone oil and deuterium dioxide immersion
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1600 nm 激发的体内深层皮肤多光子显微镜：硅油和二氧化氘浸泡的比较研究
DOI：
10.1002/jbio.202100076
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发表时间：
2021
影响因子：
2.8
通讯作者：
Qiu Ping
中科院分区：
文献类型：
--
作者：
Wang Ke;Pan Yi;Tong Shen;Chen Xinlin;Lu Yuan;Qiu Ping
在 1700 nm 窗口激发的多光子显微镜 (MPM) 能够穿透生物组织（尤其是大脑）的深层组织。皮肤的 MPM 也可能受益于这种深度渗透能力。皮肤是具有不同折射率（从 1.34 到 1.5）的层状结构。因此，在使用高数值孔径物镜成像时应选择合适的浸没介质。为了为皮肤 MPM 浸没介质的选择提供指南，我们在这里展示了使用硅油和二氧化氘 (D2O) 浸没在体内以 1600 nm 激发的深层皮肤 MPM 的比较实验研究。我们专门描述了成像深度、信号水平和空间分辨率的特征。我们的结果表明，两种浸没介质在成像深度和空间分辨率方面具有相似的性能，而硅油浸没的信号水平稍好一些。我们还证明，将荧光珠局部注射到皮肤中是体内空间分辨率表征的可行技术。
Multiphoton microscopy (MPM) excited at the 1700‐nm window has enabled deep‐tissue penetration in biological tissue, especially brain. MPM of skin may also benefit from this deep‐penetration capability. Skin is a layered structure with varying refractive index (from 1.34 to 1.5). Consequently, proper immersion medium should be selected when imaging with high numerical aperture objective lens. To provide guidelines for immersion medium selection for skin MPM, here we demonstrate comparative experimental investigation of deep‐skin MPM excited at 1600 nm in vivo, using both silicone oil and deuterium dioxide (D2O) immersion. We specifically characterize imaging depths, signal levels and spatial resolution. Our results show that both immersion media give similar performance in imaging depth and spatial resolution, while signal levels are slightly better with silicone oil immersion. We also demonstrate that local injection of fluorescent beads into the skin is a viable technique for spatial resolution characterization in vivo.