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首页> 外文期刊>Biophysical Journal >Noninvasive assessment of collagen gel microstructure and mechanics using multiphoton Microscopy
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Noninvasive assessment of collagen gel microstructure and mechanics using multiphoton Microscopy

机译：使用多光子显微镜对胶原蛋白凝胶的微观结构和力学进行无创评估

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Multiphoton microscopy of collagen hydrogels produces second harmonic generation (SHG) and two-photon fluorescence (TPF) images, which can be used to noninvasively study gel microstructure at depth (similar to 1 mm). The microstructure is also a primary determinate of the mechanical properties of the gel; thus, we hypothesized that bulk optical properties (i.e., SHG and TPF) could be used to predict bulk mechanical properties of collagen hydrogels. We utilized polymerization temperature (4-37 degrees C) and glutaraldehyde to manipulate collagen hydrogel fiber diameter, space-filling properties, and cross-link density. Multiphoton microscopy and scanning electron microscopy reveal that as polymerization temperature decreases (37-4 degrees C) fiber diameter and pore size increase, whereas hydrogel storage modulus (G', from 23 +/- 63 Pa to 0.28 +/- 0.16 Pa, respectively, mean +/- SE) and mean SHG decrease (minimal change in TPF). In contrast, glutaraldehyde significantly increases the mean TPF signal (without impacting the SHG signal) and the storage modulus (16 +/- 3.5 Pa before to 138 +/- 40 Pa after cross-linking, mean +/- SD). We conclude that SHG and TPF can characterize differential microscopic features of the collagen hydrogel that are strongly correlated with bulk mechanical properties. Thus, optical imaging may be a useful noninvasive tool to assess tissue mechanics.
机译：胶原蛋白水凝胶的多光子显微镜产生二次谐波（SHG）和双光子荧光（TPF）图像，可用于无创研究深度（约1 mm）的凝胶微观结构。微观结构也是决定凝胶机械性能的主要因素。因此，我们假设大量的光学性能（即SHG和TPF）可用于预测胶原水凝胶的整体机械性能。我们利用聚合温度（4-37摄氏度）和戊二醛来控制胶原蛋白水凝胶纤维的直径，空间填充特性和交联密度。多光子显微镜和扫描电子显微镜显示，随着聚合温度的降低（37-4摄氏度），纤维直径和孔径增加，而水凝胶的储能模量（G'）分别从23 +/- 63 Pa增至0.28 +/- 0.16 Pa ，平均+/- SE）和平均SHG降低（TPF的最小变化）。相反，戊二醛显着增加了平均TPF信号（不影响SHG信号）和储能模量（交联前为16 +/- 3.5 Pa，交联后为138 +/- 40 Pa，均值+/- SD）。我们得出的结论是，SHG和TPF可以表征胶原蛋白水凝胶的不同微观特征，这些特征与整体机械性能密切相关。因此，光学成像可能是评估组织力学的有用的非侵入性工具。

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