PDF(Pubmed)
Abstract：
Given the pivotal role of neuronal populations in various biological processes, assessing their collective output is crucial for understanding the nervous system\'s complex functions. Building on our prior development of a spiral scanning mechanism for the rapid acquisition of Raman spectra from single cells and incorporating machine learning for label-free evaluation of cell states, we investigated whether the Paint Raman Express Spectroscopy System (PRESS) can assess neuronal activities. We tested this hypothesis by examining the chemical responses of glutamatergic neurons as individual neurons and autonomic neuron ganglia as neuronal populations derived from human-induced pluripotent stem cells. The PRESS successfully acquired Raman spectra from both individual neurons and ganglia within a few seconds, achieving a signal-to-noise ratio sufficient for detailed analysis. To evaluate the ligand responsiveness of the induced neurons and ganglia, the Raman spectra were subjected to principal component and partial least squares discriminant analyses. The PRESS detected neuronal activity in response to glutamate and nicotine, which were absent in the absence of calcium. Additionally, the PRESS induced dose-dependent neuronal activity changes. These findings underscore the capability of the PRESS to assess individual neuronal activity and elucidate neuronal population dynamics and pharmacological responses, heralding new opportunities for drug discovery and regenerative medicine advancement.
摘要：
鉴于神经元群体在各种生物过程中的关键作用，评估他们的集体输出对于理解神经系统的复杂功能至关重要。基于我们先前开发的螺旋扫描机制，用于从单细胞快速获取拉曼光谱，并结合机器学习用于无标签评估细胞状态，我们调查了PaintRamanExpress光谱系统（PRESS）是否可以评估神经元活动。我们通过检查谷氨酸能神经元作为单个神经元和自主神经神经节作为源自人诱导的多能干细胞的神经元群体的化学反应来检验这一假设。PRESS在几秒钟内成功获得了单个神经元和神经节的拉曼光谱，实现足够详细分析的信噪比。为了评估诱导神经元和神经节的配体反应性，拉曼光谱进行了主成分和偏最小二乘判别分析。PRESS检测到神经元对谷氨酸和尼古丁的反应，在没有钙的情况下不存在。此外,PRESS诱导的剂量依赖性神经元活动变化。这些发现强调了PRESS评估个体神经元活动和阐明神经元种群动态和药理学反应的能力。预示着药物发现和再生医学发展的新机会。
