Mass Spectrometry Measurement of Single Suspended Cells Using a Combined Cell Manipulation System and a Single-Probe Device

Standke, Shawna J; Colby, Devon H.; Bensen, Ryan C.; Burgett, Anthony W. G.; Yang, Zhibo

doi:10.1021/acs.analchem.8b05774

Cited by 51 publications

(55 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The need to analyze large numbers of samples in single cell approaches means that robust quality control procedures need to be implemented to ensure that all the generated data is comparable. Although some studies have measured lipids in single cells (Ellis et al, 2012;Neumann et al, 2019;Thiele et al, 2019;Standke et al, 2019) and some have measured in large numbers of cells (Neumann et al, 2019), these approaches have incorporated limited quality control procedures to ensure data consistency across sample sets. In this study we also included a pooled quality control corresponding to unsorted cells, which was added to each plate and enabled us to correct for inter-plate variability (Figure 3).…”

Section: Quality Controlmentioning

confidence: 99%

“…However, without imaging all samples lack cell-type specificity and could not guarantee that each sample contained a single cell and not clusters of cells, leading to wide divergence in the number of lipids measured per sample (Neumann et al, 2019). Most single cell mass spectrometry platforms have focused on analyzing immobilized cells; however, Standke et al (2019) developed an integrated cell manipulation platform that enables single cells to be analyzed from solutions, such as bodily fluids, with minimal sample preparation. More complex derivatization approaches have also been described.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Development and Application of High-Throughput Single Cell Lipid Profiling: A Study of SNCA-A53T Human Dopamine Neurons

et al. 2020

View full text Add to dashboard Cite

Advances in single cell genomics and transcriptomics have shown that at tissue level there is complex cellular heterogeneity. To understand the effect of this inter-cell heterogeneity on metabolism it is essential to develop a single cell lipid profiling approach that allows the measurement of lipids in large numbers of single cells from a population. This will provide a functional readout of cell activity and membrane structure. Using liquid extraction surface analysis coupled with high-resolution mass spectrometry we have developed a high-throughput method for untargeted single cell lipid profiling. This technological advance highlighted the importance of cellular heterogeneity in the functional metabolism of individual human dopamine neurons, suggesting that A53T alpha-synuclein (SNCA) mutant neurons have impaired membrane function. These results demonstrate that this single cell lipid profiling platform can provide robust data that will expand the frontiers in biomedical research.

show abstract

Section: Quality Controlmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development and Application of High-Throughput Single Cell Lipid Profiling: A Study of SNCA-A53T Human Dopamine Neurons

et al. 2020

View full text Add to dashboard Cite

show abstract

“…In a time when single-cell techniques are blooming in genomics, epigenomics, transcriptomics, and proteomics [64,65], single-cell metabolomics is still in its infancy [66]. Nevertheless, recent studies have demonstrated the feasibility of different techniques for profiling the metabolome at the single-cell level from both cell cultures [67,68] and freshly resected tissues [69]. Among these techniques, mass spectrometry imaging (MSI) spatial metabolomics [70] is perhaps the most widely applicable, with the proven ability to characterize both metabolite levels and isotopic-labeling patterns at subcellular (and even suborganelle) resolutions [71].…”

Section: Box 3 Interpreting In Vivo Tracer Measurements (1): Pathwaymentioning

confidence: 99%

Stable Isotopes for Tracing Mammalian-Cell Metabolism In Vivo

Fernández-García

Altea-Manzano

Pranzini

et al. 2020

Trends in Biochemical Sciences

View full text Add to dashboard Cite

Metabolism is at the cornerstone of all cellular functions and mounting evidence of its deregulation in different diseases emphasizes the importance of a comprehensive understanding of metabolic regulation at the whole-organism level. Stable-isotope measurements are a powerful tool for probing cellular metabolism and, as a result, are increasingly used to study metabolism in in vivo settings. The additional complexity of in vivo metabolic measurements requires paying special attention to experimental design and data interpretation. Here, we review recent work where in vivo stable-isotope measurements have been used to address relevant biological questions within an in vivo context, summarize different experimental and data interpretation approaches and their limitations, and discuss future opportunities in the field. Metabolism: A Central Node for Cellular Processes Metabolism can be seen as the engine of the cell, providing energy, redox cofactors, and building blocks for cell maintenance, growth, and renewal, as well as playing a key role in modulating cell signaling [1,2]. To orchestrate all these functions, metabolism consists of a complex network of genes, enzymes, and metabolites, steadily modulated in response to different stimuli [3-5]. Defining the mechanisms at the basis of this regulation and understanding their physiological role is among the most important pursuits in biological and medical research [6], particularly since the realization that many pathologies are driven by metabolic deregulations [7]. An in-depth understanding of metabolic pathways in vivo and how these are deregulated in different diseases is thus fundamental for the discovery of new therapeutic targets and clinical biomarkers, enabling the development of more robust diagnosis approaches and personalized treatments, eventually improving the overall outcome for patients [8]. In this context, stable-isotope tracers (see Glossary) have become a standard for probing cellular metabolism [9] and their increasing implementation in in vivo settings has revolutionized our current understanding of mammalian metabolism in health and disease, unraveling novel regulatory principles at both the cellular and whole-organism level [10,11]. In this review, we summarize the latest advances in in vivo metabolic measurements using stable-isotope tracers, highlighting the importance of systems-level integrative approaches and careful experimental design, pointing out open challenges and opportunities for advancing the field, and outlining strategies and potential pitfalls when interpreting these measurements (Figure 1). Tracer-Based Methods for Measuring Cellular Metabolism In Vivo Many complementary methods exist to study metabolism in vivo and, consequently, the selection of a specific approach (or set of approaches) depends largely on the biological question being addressed. Nontracer-based methods, such as assessing bioenergetics by measuring dynamic changes in oxygen consumption [12] or evaluating changes in metabolite levels via metabolomics [13],...

show abstract

“…The ability to probe cellular heterogeneity may lead to better disease diagnosis, prognosis and treatment [1][2][3][4][5] by enabling the differentiation of subpopulations of disease-causing cells, the discovery of biochemical pathways and mechanisms, and the monitoring of the efficacy of therapeutic intervention. Technologies for performing molecular and chemical analyses at the single-cell level include fluorescence-or mass-spectrometry-based techniques [6][7][8][9][10][11] coupled with flow cytometry, [12][13][14] capillary electrophoresis [15][16][17] or optical microscopy, [18][19][20] in which the biomolecules of interest are labelled. They require complex instruments, centralized laboratory and personnel with expertise, and are often costly and time-consuming.…”

Section: Introductionmentioning

confidence: 99%