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Open AccessJournal ArticleDOI

Advances in Imaging Secondary Ion Mass Spectrometry for Biological Samples

TLDR
This work addresses the key issue of sample preparation, because mass spectrometry is performed in high vacuum, it is essential to preserve the lateral organization of the sample while removing bulk water, and this has been a major barrier for applications to biological systems.
Abstract
Imaging mass spectrometry combines the power of mass spectrometry to identify complex molecules based on mass with sample imaging. Recent advances in secondary ion mass spectrometry have improved sensitivity and spatial resolution, so that these methods have the potential to bridge between high-resolution structures obtained by X-ray crystallography and cyro-electron microscopy and ultrastructure visualized by conventional light microscopy. Following background information on the method and instrumentation, we address the key issue of sample preparation. Because mass spectrometry is performed in high vacuum, it is essential to preserve the lateral organization of the sample while removing bulk water, and this has been a major barrier for applications to biological systems. Recent applications of imaging mass spectrometry to cell biology, microbial communities, and biosynthetic pathways are summarized briefly, and studies of biological membrane organization are described in greater depth.

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Citations
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Microbial syntrophy: interaction for the common good

TL;DR: The principles of classical and non-classical syntrophy are explained and biochemical fundamentals that allow microorganism to survive under a range of environmental conditions and to drive important biogeochemical processes are presented.
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In Situ Imaging of Metals in Cells and Tissues

TL;DR: The aim of this review is to provide an overview of the most recent achievements in trace metal imaging while at the same time also offering a historical perspective of this rapidly evolving research field.
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Fluorescent Probes for Lipid Rafts: From Model Membranes to Living Cells

TL;DR: An urgent need to develop new probes, specifically adapted for cell plasma membranes and compatible with modern fluorescence microscopy techniques to push the understanding of membrane microdomains forward is highlighted.
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Mass spectrometry imaging with high resolution in mass and space

TL;DR: The high mass resolution and mass accuracy proved to be essential for specific image generation and reliable identification of analytes in tissue samples and has the potential to supplement classical histochemical protocols and to provide new insights about molecular processes on the cellular level.
Journal ArticleDOI

Imaging mass spectrometry in microbiology.

TL;DR: The wide range of imaging mass spectrometry techniques that are available to microbiologists are explored and the unique applications of these tools to microbiology with respect to the types of samples to be investigated are described.
References
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Journal ArticleDOI

Imaging intracellular fluorescent proteins at nanometer resolution.

TL;DR: This work introduced a method for optically imaging intracellular proteins at nanometer spatial resolution and used this method to image specific target proteins in thin sections of lysosomes and mitochondria and in fixed whole cells to image retroviral protein Gag at the plasma membrane.
Journal ArticleDOI

Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy

TL;DR: A new type of scanning fluorescence microscope capable of resolving 35 nm in the far field is proposed, overcome the diffraction resolution limit by employing stimulated emission to inhibit the fluorescence process in the outer regions of the excitation point-spread function.
Journal Article

Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy

TL;DR: In this paper, the authors proposed a new type of scanning fluorescence microscope capable of resolving 35 nm in the far field by employing stimulated emission to inhibit the fluorescence process in the outer regions of the excitation point spread function.
Journal ArticleDOI

Far-Field Optical Nanoscopy

TL;DR: Initial applications indicate that emergent far-field optical nanoscopy will have a strong impact in the life sciences and in other areas benefiting from nanoscale visualization.
Journal ArticleDOI

The fluorescent toolbox for assessing protein location and function

TL;DR: The focus is on protein detection in live versus fixed cells: determination of protein expression, localization, activity state, and the possibility for combination of fluorescent light microscopy with electron microscopy.
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