Neuropeptides are an important class of endogenous peptides in the nervous system that regulate physiological functions such as feeding, glucose homeostasis, pain, memory, reproduction, and many others. In order to understand the functional role of neuropeptides in diseases or disorders, studies investigating their dysregulation in terms of changes in abundance and localization must be carried out. As multiple neuropeptides are believed to play a functional role in each physiological process, techniques capable of global profiling multiple neuropeptides simultaneously are desired. Mass spectrometry is well-suited for this goal due to its ability to perform untargeted measurements without prior comprehensive knowledge of the analytes of interest. Mass spectrometry imaging (MSI) is particularly useful because it has the capability to image a large variety of peptides in a single experiment without labeling. Like all analytical techniques, careful sample preparation is critical to successful MSI analysis. The first half of this review focuses on recent developments in MSI sample preparation and instrumentation for analyzing neuropeptides and other biomolecules in which the sample preparation technique may be directly applicable for neuropeptide analysis. The benefit offered by incorporating these techniques is shown as improvement in a number of observable neuropeptides, enhanced signal to noise, increased spatial resolution, or a combination of these aspects. The second half of this review focuses on recent biological discoveries about neuropeptides resulting from these improvements in MSI analysis. The recent progress in neuropeptide detection and analysis methods, including the incorporation of various tissue washes, matrices, instruments, ionization sources, and computation approaches combined with the advancements in understanding neuropeptide function in a variety of model organisms, indicates the potential for the utilization of MSI analysis of neuropeptides in clinical settings.

神经肽是神经系统中重要的一类内源性肽，它调节生理功能，例如进食，葡萄糖稳态，疼痛，记忆，繁殖等。为了了解神经肽在疾病或失调中的功能作用，必须进行研究其在丰度和定位变化方面的失调的研究。由于相信多种神经肽在每个生理过程中都起着功能性作用，因此需要能够同时全局剖析多种神经肽的技术。质谱由于无需事先全面了解目标分析物即可执行无目标测量的能力，因此非常适合该目标。质谱成像（MSI）特别有用，因为它具有在单个实验中无需标记即可成像多种肽的能力。像所有分析技术一样，精心准备样品对于成功进行MSI分析至关重要。这篇综述的前半部分着重于MSI样品制备和用于分析神经肽和其他生物分子的仪器的最新进展，其中样品制备技术可能直接适用于神经肽分析。合并这些技术所提供的好处显示为可观察到的神经肽数量有所改善，信噪比提高，空间分辨率提高或这些方面的组合。这篇综述的后半部分着重介绍了MSI分析中的这些改进导致的有关神经肽的最新生物学发现。神经肽检测和分析方法的最新进展，包括各种组织洗涤液，基质，仪器，电离源和计算方法的结合，以及对多种模型生物中神经肽功能的理解的进步，表明利用神经肽的潜力。在临床环境中对神经肽进行MSI分析。