The impact of cell fixation on coherent anti-stokes Raman scattering signal intensity in neuronal and glial cell lines
The impact of cell fixation on coherent anti-stokes Raman scattering signal intensity in neuronal and glial cell lines
复制标题
细胞固定对神经元和神经胶质细胞系相干反斯托克斯拉曼散射信号强度的影响
DOI：
10.1002/jbio.201800203
复制
发表时间：
2018
影响因子：
2.8
通讯作者：
Qu Junle
中科院分区：
文献类型：
--
作者：
Levchenko Svitlana M.;Peng Xiao;Liu Liwei;Qu Junle
许多研究需要样品固定，目的是将细胞保存在生理状态下，使形态和细胞内分子含量的变化最小。样品固定可能会严重扭曲实验数据，这使得数据解释过程更具挑战性。这对于脂质相关疾病的研究尤其重要，因为细胞的生化和形态特征需要得到很好的保存，以便进行准确的数据分析。本研究研究了甲醛和乙醇（EtOH）固定剂对神经元和神经胶质细胞主要细胞区室中蛋白质和脂质的相干反斯托克斯拉曼散射（汽车）信号的影响。我们发现，这两种固定剂诱导蛋白质和脂质信号在所研究的细胞系的改变。此外，样品保存方法对汽车信号的影响在细胞系之间不同。例如，我们的数据显示，EtOH固定诱导神经元细胞核仁中蛋白质的汽车信号增加约45%，神经胶质细胞中汽车信号减少约35%。结果表明，醛固定是一种较好的方法，用于保存神经元和神经胶质细胞的汽车成像之前，因为它较少影响汽车信号和细胞内分布的蛋白质和脂质。
A number of studies require sample fixation, aimed to preserve cells in a physiological state with minimal changes of morphology and intracellular molecular content. Sample fixation may significantly distort experimental data, which makes the data interpretation process more challenging. It is particularly important for study of lipid‐related diseases, where the biochemical and morphological characteristics of the cells need to be well preserved for an accurate data analysis. This study investigates the effects of formaldehyde and ethanol (EtOH) fixatives on coherent anti‐stokes Raman scattering (CARS) signal of proteins and lipids in major cellular compartments of neuronal and glial cells. We found that both fixatives induce alteration of proteins and lipids signal in studied cell lines. Furthermore, the impact of sample preservation methods on CARS signal varies between cell lines. For instance, our data reveals that EtOH fixation induces ~45% increase of CARS signal of proteins in the nucleolus of neuronal cells and ~35% decrease of CARS signal in glial cells. The results indicate that aldehyde fixation is a preferable method for preservation of neuronal and glial cells prior to CARS imaging, as it less affects both CARS signal and intracellular distribution of proteins and lipids.