Establishment of a novel platform of Raman microscope for diagnosis of hydrogel-generated cancer stem cells.

在本财政年度期间，研究了PAMPS凝胶对成肌细胞模型的作用。发现聚（2-丙烯酰胺基-2-甲基-1-丙磺酸）水凝胶对诱导干性标志物（如Sox 2、Nanog和Oct 3/4）的升高具有显著影响。这种干性标志物的升高与细胞形态的变化有关，这意味着具有较高表达水平的细胞往往更圆和更小，而具有低干性标志物表达的细胞往往更大和更平。用高速狭缝扫描拉曼显微镜拍摄了上述水凝胶-细胞相互作用体系的第一张拉曼图像。测量的质量用不同的度量进行，例如信噪比、拉普拉斯算子和非均匀线照明量化。为了利用水凝胶基质上的高光谱拉曼图像，最终确定了两种算法，可以比较来自不同拉曼设备和不同测量日期的数据。一种算法用于单细胞测量。另一个是用于3D集落形成测量。

During this fiscal year was investigated the action of PAMPS gel on myoblast cell models. It was found that Poly (2-acrylamido-2- methyl-1-propanesulphonic acid) hydrogel have a significant impact to induce an elevation of stemness markers such as Sox2, Nanog, and Oct3/4. This elevation of stemness markers have been linked to a change of cell morphology, meaning that cell that have a higher expression level tend to be more round and small whereas the one that have a low expression of stemness markers tend to be bigger and flat. This interesting fact showed that by stainingThe first Raman images of the mentioned hydrogel-cell interaction system was performed with a high-speed slit-scanning Raman microscope. The quality of the measurement was performed with different metric such as signal-to-noise ratio, Laplacian operator and non-homogenous line illumination quantification. As a result the measurement protocol for this complex measurement provide high-quality data.To exploit Hyperspectral Raman images on hydrogel substrate, two algorithms that allow to compare Data from different Raman devices and different dates of measurement was finalized. One algorithm is for single cell measurement. Another one is for 3D colony formation measurement.