Single Cell Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging

Schober, Yvonne; Guenther, Sabine; Spengler, Bernhard; Römpp, Andreas

doi:10.1021/ac301337h

Cited by 179 publications

(173 citation statements)

References 30 publications

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“…Furthermore, recent applications of AP-MALDI using tightly focused laser beams (spot size ~5-7 µm) has allowed the lipid composition of single cells, including HeLa cells [101] when only a few unique lipids were present at a given sampling spot. Glycerol adducts have also been observed for GL analysis in negative ion mode [114], and are attributed to the use of an elevated pressure ionization source and associated collisional cooling of the adducts.…”

Section: Atmospheric Pressure Maldi (Ap-maldi)mentioning

confidence: 99%

Surface analysis of lipids by mass spectrometry: More than just imaging

Ellis

Brown

Panhuis

et al. 2013

Progress in Lipid Research

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Mass spectrometry is now an indispensable tool for lipid analysis and is arguably the driving force in the renaissance of lipid research. In its various forms, mass spectrometry is uniquely capable of resolving the extensive compositional and structural diversity of lipids in biological systems. Furthermore, it provides the ability to accurately quantify molecular-level changes in lipid populations associated with changes in metabolism and environment; bringing lipid science to the "omics" age. The recent explosion of mass spectrometry-based surface analysis techniques is fuelling further expansion of the lipidomics field. This is evidenced by the numerous papers published on the subject of mass spectrometric imaging of lipids in recent years. While imaging mass spectrometry provides new and exciting possibilities, it is but one of the many opportunities direct surface analysis offers the lipid researcher. In this review we describe the current state-ofthe-art in the direct surface analysis of lipids with a focus on tissue sections, intact cells and thin-layer chromatography substrates. The suitability of these different approaches towards analysis of the major lipid classes along with their current and potential applications in the field of lipid analysis are evaluated. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 2 AbstractMass spectrometry is now an indispensable tool for lipid analysis and is arguably the driving force in the renaissance of lipid research. In its various forms, mass spectrometry is uniquely capable of resolving the extensive compositional and structural diversity of lipids in biological systems. Furthermore, it provides the ability to accurately quantify molecular-level changes in lipid populations associated with changes in metabolism and environment;bringing lipid science to the "omics" age. The recent explosion of mass spectrometry-based surface analysis techniques is fuelling further expansion of the lipidomics field. This is evidenced by the numerous papers published on the subject of mass spectrometric imaging of lipids in recent years. While imaging mass spectrometry provides new and exciting possibilities, it is but one of the many opportunities direct surface analysis offers the lipid researcher. In this review we describe the current state-of-the-art in the direct surface analysis of lipids with a focus on tissue sections, intact cells and thin-layer chromatography substrates.The suitability of these different approaches towards analysis of the major lipid classes along with their current and potential applications in the field of lip...

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Section: Atmospheric Pressure Maldi (Ap-maldi)mentioning

confidence: 99%

Surface analysis of lipids by mass spectrometry: More than just imaging

Ellis

Brown

Panhuis

et al. 2013

Progress in Lipid Research

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“…Matrix-assisted laser desorption ionization (MALDI) imaging mass spectrometry is the method of choice for the analysis of tissues (13), and new laser desorption sources render it possible to image the distribution of small molecules with single-cell resolution (14,15). Together with its high-throughput capabilities, single-cell MALDI-MS can give Screening of Chlamydomonas reinhardtii populations with single-cell resolution by using a high-throughput microscale sample preparation for matrix-assisted laser desorption ionization mass spectrometry.…”

mentioning

confidence: 99%

Screening of Chlamydomonas reinhardtii Populations with Single-Cell Resolution by Using a High-Throughput Microscale Sample Preparation for Matrix-Assisted Laser Desorption Ionization Mass Spectrometry

Krismer

Sobek

Steinhoff

et al. 2015

Appl Environ Microbiol

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bThe consequences of cellular heterogeneity, such as biocide persistence, can only be tackled by studying each individual in a cell population. Fluorescent tags provide tools for the high-throughput analysis of genomes, RNA transcripts, or proteins on the single-cell level. However, the analysis of lower-molecular-weight compounds that elude tagging is still a great challenge. Here, we describe a novel high-throughput microscale sample preparation technique for single cells that allows a mass spectrum to be obtained for each individual cell within a microbial population. The approach presented includes spotting Chlamydomonas reinhardtii cells, using a noncontact microarrayer, onto a specialized slide and controlled lysis of cells separated on the slide. Throughout the sample preparation, analytes were traced and individual steps optimized using autofluorescence detection of chlorophyll. The lysates of isolated cells are subjected to a direct, label-free analysis using matrix-assisted laser desorption ionization mass spectrometry. Thus, we were able to differentiate individual cells of two Chlamydomonas reinhardtii strains based on single-cell mass spectra. Furthermore, we showed that only population profiles with real single-cell resolution render a nondistorted picture of the phenotypes contained in a population. Heterogeneity plays a pivotal role in the emergence of tolerance, persistence, and resistance toward biocides in microbial populations (1, 2). Also, microbial populations show highly complex interactions, e.g., in the competition for nutrients or in the colonization of new habitats (3). In recent years, newly developed tools for single-cell analysis have greatly extended our understanding of biological variation in microbial populations and its underlying mechanisms (4). These tools allow genome sequencing (5) or follow transcription as well as protein synthesis on the single-cell level (6). Since these techniques give a much higher resolution when observing processes in given cell populations, they permit insight into inter-and intracellular processes and the underlying mechanisms. However, when it comes to highthroughput measurements of small molecules, very few methods are currently known (7).The singular qualities of individual cells can only be fully appreciated within the context of the population. Therefore, one of the most important features for single-cell methods to be useful in biological applications is high-throughput capability. One of the most successful high-throughput approaches to characterize heterogeneities in microbial populations is flow cytometry (8). It has the benefits of high sensitivity and a high linear dynamic range of fluorescence tagging and optical detection. However, the method is strongly limited in parallelization, since excitation and emission bands overlap. Mass cytometry, on the other hand, which is a new approach that can be coupled to flow cytometry, uses antibodies tagged with rare earth metals (9). With mass spectrometric detection, over 40 features can be meas...

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“…Schober et al was the first to develop single-cell MALDI-IMS analysis using the HeLa cell line [27]; however, no report has examined comparative lipid profiling by single-cell MALDI-IMS analysis. Several reports have been published on comparative analysis of single cells using secondary ion mass spectrometry (SIMS), and these reports have demonstrated fatty acid profiles in the single-cell level using clinical samples such as breast cancer stem cell and MM cell [28,29].…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, single-cell MALDI-IMS analysis is a useful method for demonstrating lipid profiles and detecting rare target populations in human clinical samples. We suppose that the following methodological advances in the field of IMS made it possible to analyze such small objects: (1) the combined use of a microscope together with IMS and an improved laser ablation area to increase the spatial resolution up to 5 μm [27,31]; (2) the use of DHAP as a matrix for improved sensitivity of phospholipid detection by MALDI-IMS [21]; and (3) the sample preparation strategies such as the introduction of glutaraldehyde for cell fixation in IMS analysis [20].…”

Section: Discussionmentioning

confidence: 99%

Decreased level of phosphatidylcholine (16:0/20:4) in multiple myeloma cells compared to plasma cells: a single-cell MALDI–IMS approach

et al. 2015

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Lipid metabolic changes under diseased conditions, particularly in solid tumors, are attracting increased attention. However, in non-solid tumors, including most hematopoietic tumors, lipid analyses are scarce. Multiple myeloma (MM) is a plasma cell disorder arising from bone marrow, and the lipid status of MM cells has not been reported yet. In this study, we analyzed flow cytometry-sorted single MM cells and normal plasma cells (NPCs) using matrix-assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS), a two-dimensional label-free mass spectrometry technique for biomolecular analysis, to obtain specific lipid information. We isolated 1.31-5.77% of MM cells and 0.03-0.24% of NPCs using fluorescence-activated cell sorting (FACS). Analysis of purified cells using MALDI-IMS at the single-cell level revealed that the peak intensity and ion signals of phosphatidylcholine [PC (16:0/20:4) + H](+) at m/z 782.5 were significantly decreased in MM cells compared to NPCs. By examining particular cell populations rather than cell mixtures, our method can become a suitable tool for the analysis of rare cell populations at the single-cell level and advance the understanding of MM progression.

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Single Cell Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging

Cited by 179 publications

References 30 publications

Surface analysis of lipids by mass spectrometry: More than just imaging

Surface analysis of lipids by mass spectrometry: More than just imaging

Screening of Chlamydomonas reinhardtii Populations with Single-Cell Resolution by Using a High-Throughput Microscale Sample Preparation for Matrix-Assisted Laser Desorption Ionization Mass Spectrometry

Decreased level of phosphatidylcholine (16:0/20:4) in multiple myeloma cells compared to plasma cells: a single-cell MALDI–IMS approach

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