乙醇胁迫酵母凋亡过程的单细胞喇曼光谱

Apoptosis Process under High Concentrations of Ethanol Stress at a Single Cell Level with Raman Spectroscopy

在群体细胞水平和单个酵母细胞水平上,应用光镊喇曼光谱技术对高浓度乙醇胁迫过程中细胞内生物大分子物质的动态变化进行实时监测.基于两种水平上的研究结果显示:在高浓度乙醇胁迫下,归属于核酸的喇曼峰(721,1 083,1 301cm-1)、归属于蛋白质的喇曼峰(721,858,1 001,1 301,1 445,1 608,1 657cm-1)和归属于脂类(1 083,1 301,1 445cm-1)喇曼峰的峰强度都随处理时间的延长而显著降低,说明了高浓度乙醇诱导酵母凋亡过程中核酸、蛋白质、脂类等物质的含量是逐渐降低的.结合单因素分析法与重复测量分析法发现,群体细胞与单个细胞光谱的变化有明显差异性,反映基于群体细胞的研究,个体细胞的信息被平均光谱信息所掩盖,无法体现个体间所存在的差异.应用激光镊子喇曼光谱技术基于单个细胞水平上的研究能更直接、真实地反映高浓度乙醇胁迫下细胞内生物大分子变化的动态信息.

At group cells level and a single yeast level, laser tweezers Raman spectroscopy was used to monitor the dynamic of the intracellular biological macromolecular in real-time during the apoptosis process of yeast cells stressed with high concentrations of ethanol. The results show that the intensities of Raman peaks of nucleic acids （721, 1 083, 1 301 cm^-1）, proteins （721, 858, 1 001, 1 301, 1 445, 1 608, 1 657 cm^-1） and lipids （1 083, 1 301, 1 445 cm^-1） decreased significantly along with the extension of treatment time, indicates that the contents of nucleic acids, proteins and lipids are reduced gradually while the yeast cells are undergoing apoptosis induced by high concentrations of ethanol. However, the results of single factor analysis and repeated measures analysis show that the changes of the intensities of Raman peaks between the group cells and the single cells are difference significantly, implies that the heterogeneities of the single cells are covered by the average spectroscopy of the population cells. The results display that laser tweezer Raman spectroscopy can be used to directly and truthfully detect the kinetic apoptosis process under high concentrations of ethanol stress at the single cell level and probe cellular heterogeneity.