Vitamin E Imaging and Localization in the Neuronal Membrane

Monroe, Eric B.; Jurchen, John C.; Lee, Jinju; Rubakhin, Stanislav S.; Sweedler, Jonathan V.

doi:10.1021/ja051223y

Cited by 124 publications

(159 citation statements)

References 23 publications

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“…For example, Monroe et al showed that vitamin E (m/z 430) was localized at the junction of the cell soma and neurite in single isolated neurons of the California sea slug. 38 Lipid localization in cell membranes has also been observed in both glial and neuroblastoma cells. 37,39 C 60 + primary ion sources have generally not been used to image cells because they are typically focused to only ∼1 µm.…”

Section: Biological Imaging Of Tissues and Cellsmentioning

confidence: 99%

Why Is SIMS Underused in Chemical and Biological Analysis? Challenges and Opportunities

Walker

2008

Anal. Chem.

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Improvements have led to many developments in SIMS, including better 2D MS imaging, the ability to perform molecular depth profiling, and the development of 3D MS imaging. (To listen to a podcast about this feature, please go to the Analytical Chemistry website at pubs.acs.org/ac.)Imaging MS has the unique ability to acquire the spatial distribution of a wide range of atoms and molecules, including polymers, pharmaceuticals, lipids, proteins, and semiconductors, without the use of labels such as fluorescent tags. Secondary ion MS (SIMS) imaging has advantages over MALDI imaging: it has submicrometer resolution and requires no sample preparation. Although MALDI imaging has been very successful, particularly in biological applications, 1 and has an increasing number of practitioners, SIMS imaging does not seem to be receiving the same amount of attention. In SIMS, ion beams are used to desorb molecules from surfaces; the mass of the molecules is then measured by MS. It was one of the first techniques developed for detecting organic molecules that were not amenable to electron impact ionization. 2,3It was largely overshadowed by other MS techniques, including ESI and MALDI, because in SIMS, it was difficult to desorb ions with m/z > 500, which makes the detection of large molecules very hard. In recent years, SIMS has undergone a renaissance with the advent of commercially available cluster ion sources: Au n + , Bi n x+ , SF 5 + , and C 60 x+ . [4][5][6][7][8][9][10][11] These are easy to use, reliable, and have long lifetimes (g500 µA/h). Cluster ion beams produce secondary ions with much higher efficiencies than atomic ion sources do, thereby increasing secondary ion yields and decreasing sample damage. These improvements have led to many developments in SIMS technology, including better 2D MS imaging, the ability to perform molecular depth profiling, and the development of 3D MS imaging. In this article, I will highlight some of the new applications enabled by cluster primary ion beams, particularly in biological imaging, and I will discuss the challenges of implementing SIMS more widely. (This article is not a comprehensive review of SIMS; for additional information, see Refs. 1-3 and 12-25.) IMAGE RESOLUTION AND CLUSTER ION SOURCESIn imaging MS, the image quality is determined by three different resolutions: mass, lateral, and depth. Mass resolution, m/∆m, is important because it determines the chemical specificity of the technique as well as its mass accuracy (through improved precision). For many years, commercially available TOF SIMS instruments have been able to produce m/∆m of several thousand in imaging mode.

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Section: Biological Imaging Of Tissues and Cellsmentioning

confidence: 99%

Why Is SIMS Underused in Chemical and Biological Analysis? Challenges and Opportunities

Walker

2008

Anal. Chem.

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“…This technique has already been used to image tocopherol location in cells [102] and tissues [103][104][105]. It would be fascinating if someone should now apply the tools of lipidomics to collecting data not only on the location of tocopherol, but also the phospholipid profile at the same location.…”

Section: Is All Of This Relevant For Understanding the Biological Actmentioning

confidence: 99%

The location and behavior of α‐tocopherol in membranes

Atkinson

Harroun

Wassall

et al. 2010

Molecular Nutrition Food Res

168

128

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Vitamin E (a-tocopherol) has long been recognized as the major antioxidant in biological membranes, and yet many structurally related questions persist of how the vitamin functions. For example, the very low levels of a-tocopherol reported for whole cell extracts question how this molecule can successfully protect the comparatively enormous quantities of PUFAcontaining phospholipids found in membranes that are highly susceptible to oxidative attack. The contemporary realization that membranes laterally segregate into regions of distinct lipid composition (domains), we propose, provides the answer. We hypothesize a-tocopherol partitions into domains that are enriched in polyunsaturated phospholipids, amplifying the concentration of the vitamin in the place where it is most needed. These highly disordered domains depleted in cholesterol are analogous, but organizationally antithetical, to the wellstudied lipid rafts. We review here the ideas that led to our hypothesis. Experimental evidence in support of the formation of PUFA-rich domains in model membranes is presented, focusing upon docosahexaenoic acid that is the most unsaturated fatty acid commonly found. Physical methodologies are then described to elucidate the nature of the interaction of a-tocopherol with PUFA and to establish that the vitamin and PUFA-containing phospholipids co-localize in non-raft domains.

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“…1 Numerous analytical techniques, including electrochemical detection, 2-6 separation-based methods, 7-9 fluorescence microscopy, 10-13 and mass spectrometry [14][15][16][17][18][19][20][21][22][23][24] have been used to surmount the inherent difficulties of measurements down to the μm-, pL-, and zmole-scale to study the intricate chemistry that exists within single cells.…”

Section: Introductionmentioning

confidence: 99%

Secondary Ion MS Imaging To Relatively Quantify Cholesterol in the Membranes of Individual Cells from Differentially Treated Populations

et al. 2007

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Time of flight secondary ion mass spectrometry (ToF-SIMS) is a well-established bioanalytical method for directly imaging the chemical distribution across single-cells. Here we report a protocol for the use of SIMS imaging to comparatively quantify the relative difference in lipid composition between the plasma membranes of two cells. This development enables direct comparison of the chemical effects of different drug treatments and incubation conditions in the plasma membrane at the single-cell level. Relative, quantitative ToF-SIMS imaging has been used here to compare macrophage cells treated to contain elevated levels of cholesterol with respect to control cells. Insitu fluorescence microscopy with two different membrane dyes was used to discriminate morphologically similar but differentially treated cells prior to SIMS analysis. SIMS images of fluorescently identified cells reveal that the two populations of cells have distinct outer leaflet membrane compositions with the membranes of the cholesterol-treated macrophages containing more than twice the amount of cholesterol of control macrophages. Relative quantification with SIMS to compare the chemical composition of single-cells can provide valuable information about normal biological functions, causative agents of diseases, and possible therapies for diseases.

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Vitamin E Imaging and Localization in the Neuronal Membrane

Cited by 124 publications

References 23 publications

Why Is SIMS Underused in Chemical and Biological Analysis? Challenges and Opportunities

Why Is SIMS Underused in Chemical and Biological Analysis? Challenges and Opportunities

The location and behavior of α‐tocopherol in membranes

Secondary Ion MS Imaging To Relatively Quantify Cholesterol in the Membranes of Individual Cells from Differentially Treated Populations

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