Single Cell Mass Spectrometry

Romanova, Elena V.; Rubakhin, Stanislav S.; Monroe, Eric B.; Sweedler, Jonathan V.

doi:10.1002/9783527626649.ch6

Cited by 6 publications

(7 citation statements)

References 101 publications

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“…Several studies have attempted to improve the methods for analyzing the chemical composition of a group of cells in the same tissue − or of even single cells. − Single-cell analysis in plant tissues is different from that of cultured cells, because of the plant tissues’ irregular surfaces, higher dilution of biomolecules in the cell, and strong cellulose walls. These are factors to be considered when designing a method for direct single-cell analysis in undamaged plant tissue.…”

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confidence: 99%

“…MS has been applied to metabolic analysis at the single-cell level. , High-resolution mass spectrometers, such as a hybrid linear ion trap/Orbitrap mass spectrometer, have enabled the identification of both common and uncommon compounds. , The combination of video-microscopic imaging and MS has been used to directly perform live single-cell analyses. ,,, We have extended this technique for directly characterizing live plant tissues to provide fast, versatile, and nondestructive analysis of different single cells in several types of plant tissues. This allows the direct collection of information at the cellular level in real time.…”

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confidence: 99%

“…MS has been applied to metabolic analysis at the single-cell level. 23,24 High-resolution mass spectrometers, such as a hybrid linear ion trap/Orbitrap mass spectrometer, have enabled the identification of both common and uncommon compounds. 25,26 The combination of video-microscopic imaging and MS has been used to directly perform live single-cell analyses.…”

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confidence: 99%

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In Situ Molecular Analysis of Plant Tissues by Live Single-Cell Mass Spectrometry

et al. 2012

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We report the development of a rapid, direct molecular analysis of live, single plant cells viewed under a video microscope in their natural environment. A nanoelectrospray tip was used to extract the contents of a single leaf, stem, or petal cell from Pelargonium zonale, and the samples were analyzed on an Orbitrap mass spectrometer by nanoelectrospray ionization. Around a thousand m/z peaks belonging to metabolites and other compounds in each sample were obtained and processed by using statistical tools to find the cell specific molecular peaks. Hybrid high-resolution mass spectrometry analysis was performed to confirm the structure of specific metabolites from the analyzed samples. This method is useful for identifying specific molecules in live single cells from plant tissue and will allow different cell types and stages from different sites in the plant to be compared with morphological observations.

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In Situ Molecular Analysis of Plant Tissues by Live Single-Cell Mass Spectrometry

et al. 2012

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“…In an early work on single-cell mass spectrometry, a laser microprobe mass analyzer (LAMMA) was applied to detect inorganic cations in single mycobacteria . Since then, a number of studies on single-cell analysis using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) have been published (for reviews, see, for example, refs and ). Recently, it was also shown that, by utilizing a tightly focused laser beam in matrix-free laser desorption/ionization (LDI), it is possible to visualize the distribution of naphthodianthrones and flavonoids in plant cells, and the potential of atmospheric pressure (AP) ionization MS approaches in this context has been successfully demonstrated in studies using AP-MALDI imaging for tissue analysis and a modified laser ablation electrospray ionization (LAESI) approach for single-cell analysis …”

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Multidimensional Analysis of Single Algal Cells by Integrating Microspectroscopy with Mass Spectrometry

et al. 2011

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We demonstrate a facile label-free approach for performing multidimensional chemical analysis on individual single-cell organisms by combining optical, fluorescence, and Raman microspectroscopy with matrix-free laser desorption/ionization mass spectrometry (MS). Single unicellular algae are seeded on a bare stainless steel plate and analyzed microspectroscopically. This provides information on the content and distribution of photoactive species, such as β-carotene, as well as chlorophyll and other components of the photosynthetic apparatus. Exactly the same cells are then analyzed by mass spectrometry in the negative ion mode. Phospholipid species are readily ionized by laser desorption/ionization of intact cells, without the need for an auxiliary matrix. This not only facilitates sample preparation but also preserves high spatial resolution and high sensitivity. Using this method, we were able to study the content and arrangement of proplastids and photosystem components, as well as the amounts of various phospholipid species in individual algal cells. The methodology can be used in the fundamental biological studies on these unicellular organisms, which require information on the internal structure as well as the chemical composition of individual cells.

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“…State-of-the-art MALDI-MS instrumentation can provide a spatial resolution that is sufficient for numerous imaging applications . Despite demonstration of several protocols for MALDI-MS analysis on the cellular and subcellular level (for a review on single-cell MS, see, for example, ref ), preparation of microscopic samples (such as cells or small cell ensembles) for MALDI imaging is still challenging. Issues to address encompass artifact-free sample handling, preservation of the native chemical composition, and targeting analyte molecules in specific cell compartments.…”

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Microscale MALDI Imaging of Outer-Layer Lipids in Intact Egg Chambers fromDrosophila melanogaster

Urban

Chang

et al. 2011

Anal. Chem.

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Fruit fly (Drosophila melanogaster) is a standard model organism used in genetics and molecular biology. Phospholipids are building blocks of cellular membranes, and components of a complex signaling network. Here, we present a facile method, based on matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS), for molecular imaging of phospholipid distributions in submillimeter-sized components of the fruit fly reproductive system. Individual egg chambers were deposited on a specially prepared MALDI target comprising an aluminum slide with a rough surface created by ablation with a microsecond-laser: this helped to immobilize biological specimens, remove excess of saline solution by adhesive forces, carry out microscopic observations, and facilitated distribution of the MALDI matrix. A continuous-flow ultrasound-assisted spray was used for the deposition of MALDI matrix (9-aminoacridine) onto the sample. The upper surface of the specimen was then scanned with a 355-nm solid-state laser with a preset beam focus of 10 μm to obtain negative-ion mode MALDI-MS images. Overall, this provided sufficient spatial resolution to reveal micrometer-scale gradient-like patterns of phospholipids along the anterior/posterior axis of egg chambers. Several phosphatidylinositols are seen to be segregated according to the number of unsaturated bonds, with an elevated abundance of polyunsaturated phosphatidylinositols within the oocyte compartment.

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Single Cell Mass Spectrometry

Cited by 6 publications

References 101 publications

In Situ Molecular Analysis of Plant Tissues by Live Single-Cell Mass Spectrometry

In Situ Molecular Analysis of Plant Tissues by Live Single-Cell Mass Spectrometry

Multidimensional Analysis of Single Algal Cells by Integrating Microspectroscopy with Mass Spectrometry

Microscale MALDI Imaging of Outer-Layer Lipids in Intact Egg Chambers fromDrosophila melanogaster

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