Quantitative structural markers of colorectal dysplasia in a cross sectional study of ex vivo murine tissue using label-free multiphoton microscopy.
Quantitative structural markers of colorectal dysplasia in a cross sectional study of ex vivo murine tissue using label-free multiphoton microscopy.
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使用无标记多光子显微镜对离体小鼠组织进行横断面研究中结直肠发育不良的定量结构标记。
DOI：
10.1117/12.2213069
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Muldoon,TimothyJ
中科院分区：
文献类型：
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作者：
Prieto,SandraP;Greening,GageJ;Lai,KeithK;Muldoon,TimothyJ
无标记组织的双光子激发越来越受到关注，因为多光子显微镜在内窥镜临床应用中取得了进展，例如用于监测小鼠宫颈胶原蛋白的二次谐波发生（SHG）扫描内窥镜1。我们使用C57 BL小鼠作为模型来研究胃肠结构的进展，特别是腺面积和循环。我们使用多光子显微镜成像离体无标记小鼠结肠，重点是随着时间的推移，在小鼠10至20周龄的胶原结构的变化。在结肠和肠组织内采集一系列图像，从肌层到上皮的深度间隔为20微米，最大深度为180微米。成像系统包括调谐到800 nm波长激发的双光子激光器，并且在到达光电倍增管之前用400/40带通滤波器过滤SHG发射。以每秒15帧的速度采集图像，累积200至300帧，视场为261 μ m × 261 μ m，样品处功率为40 mW。将图像系列与从相邻位置拍摄的组织病理学H&E载玻片进行比较。应用用于确定鼠腺结构之间差异的定量度量，特别是腺面积和圆形度。
Two-photon excitation of label-free tissue is of increasing interest, as advances have been made in endoscopic clinical application of multiphoton microscopy, such as second harmonic generation (SHG) scanning endoscopy used to monitor cervical collagen in mice1. We used C57BL mice as a model to investigate the progression of gastrointestinal structures, specifically glandular area and circularity. We used multiphoton microscopy to image ex-vivo label-free murine colon, focusing on the collagen structure changes over time, in mice ranging from 10 to 20 weeks of age. Series of images were acquired within the colonic and intestinal tissue at depth intervals of 20 microns from muscularis to the epithelium, up to a maximum depth of 180 microns. The imaging system comprised a two-photon laser tuned to 800nm wavelength excitation, and the SHG emission was filtered with a 400/40 bandpass filter before reaching the photomultiplier tube. Images were acquired at 15 frames per second, for 200 to 300 cumulative frames, with a field of view of 261um by 261um, and 40mW at sample. Image series were compared to histopathology H&E slides taken from adjacent locations. Quantitative metrics for determining differences between murine glandular structures were applied, specifically glandular area and circularity.