![]() ![]() ![]() (13−16) For this reason, single cell omics is increasingly applied to decipher the degree of biochemical differences between cells within a tissue to understand phenomena such as immune cell plasticity, (17) microbial resistance, (18) and cellular dysregulation. Single cell scales are also necessary to elucidate changes in cell biochemistry that occur in the early stages of a disease, for example, which might otherwise be unresolvable by conventional bulk-based measurements. Single cell omics differs from bulk omics in that analysis and/or extraction of nucleic acids, proteins, and metabolites is constrained and targeted to singular cells within a tissue or a culture, the benefit of which is that intercellular differences can be measured, allowing for the visualization of discrete populations of cells based on their physiological states ( Figure 1). The biological and bioanalytical chemistry research communities have increasingly set their sights on adapting MS-based proteomics and metabolomics to the single cell scale. (2−5) Innovations in MS have resulted in an explosion of discoveries and technological advancements within these omic fields over the last two decades (6−8) and have allowed the scientific community to provide answers to complex biological questions, including how protein expression regulates the circadian clock and cell signal transduction, (9,10) as well as enable the development of tools to artificially evolve proteins or target specific gene sequences for modification. In the fields of proteomics and metabolomics, mass spectrometry (MS) has been invaluable as these methods are ubiquitously employed for bulk characterization of proteins and metabolites extracted from homogenized tissue and cell lysates. (1) Molecular-based omics can be subcategorized into genomics, transcriptomics, proteomics, and metabolomics, all of which provide valuable information toward understanding the complete biotic state of biological systems. Omics measurements involve the identification and quantification of biomolecules with the overall aim of inferring the physiological state of an organism based on molecular type, location, and any change in abundance. Finally, using this broad literature summary, we provide a perspective on how the next decade may unfold in the area of single cell MS-based proteomics and metabolomics. In this Perspective, we showcase advancements in single cell spatial metabolomics and proteomics over the past decade and highlight important aspects related to high-throughput screening, data analysis, and more which are vital to the success of achieving proteomic and metabolomic profiling at the single cell scale. For example, reporter-based methods using heavy metal tags have allowed for targeted MS investigation of the proteome at the subcellular level, and development of technologies such as laser ablation electrospray ionization mass spectrometry (LAESI-MS) now mean that dynamic metabolomics can be performed in situ. Of these technologies, spatially resolved mass spectrometry approaches, including mass spectrometry imaging (MSI), have shown the most progress for single cell proteomics and metabolomics. Over the past decade, substantial advancements in our ability to characterize omic profiles on a single cell level have occurred, including in multiple spectroscopic and mass spectrometry (MS)-based techniques. ![]() Biological function varies greatly across populations of cells, as each single cell has a unique transcriptome, proteome, and metabolome that translates to functional differences within single species and across kingdoms. Biological systems are composed of heterogeneous populations of cells that intercommunicate to form a functional living tissue. ![]()
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