Improvement of biomolecular analysis in thin films using in-situ matrix enhanced Secondary Ion Mass Spectrometry

The Analyst ◽  
2021 ◽  
Author(s):  
Konstantin Moshkunov ◽  
Benjamin Tomasetti ◽  
Thomas Daphnis ◽  
Vincent Delmez ◽  
Kevin Vanvarenberg ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Secondary Ion Mass Spectrometry ◽  
Imaging Mass Spectrometry ◽  
Molecular Ions ◽  
Limiting Factor ◽  
Ion Mass Spectrometry ◽  
Resolution Imaging ◽  
Biomolecular Analysis ◽  
Secondary Ion

Sensitivity to molecular ions remains a limiting factor for high resolution imaging mass spectrometry of organic and biological materials. Here, we investigate a variant of matrix-enhanced secondary ion mass spectrometry...

Optics-Free Visualization of Proteins in Single Cells by Time of Flight-Secondary Ion Mass Spectrometry Coupled with Genetically Encoded Chemical Tags

2020 ◽  
Author(s):  
Feifei Jia ◽  
Jie Wang ◽  
Yanyan Zhang ◽  
Qun Luo ◽  
Luyu Qi ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Secondary Ion Mass Spectrometry ◽  
Single Cells ◽  
Time Of Flight ◽  
E Coli ◽  
Tof Sims ◽  
Ion Mass Spectrometry ◽  
Chemical Tags ◽  
Secondary Ion

<p></p><p><i>In situ</i> visualization of proteins of interest at single cell level is attractive in cell biology, molecular biology and biomedicine, which usually involves photon, electron or X-ray based imaging methods. Herein, we report an optics-free strategy that images a specific protein in single cells by time of flight-secondary ion mass spectrometry (ToF-SIMS) following genetic incorporation of fluorine-containing unnatural amino acids as a chemical tag into the protein via genetic code expansion technique. The method was developed and validated by imaging GFP in E. coli and human HeLa cancer cells, and then utilized to visualize the distribution of chemotaxis protein CheA in E. coli cells and the interaction between high mobility group box 1 protein and cisplatin damaged DNA in HeLa cells. The present work highlights the power of ToF-SIMS imaging combined with genetically encoded chemical tags for <i>in situ </i>visualization of proteins of interest as well as the interactions between proteins and drugs or drug damaged DNA in single cells.</p><p></p>

scholarly journals Triple Oxygen Isotope Measurements by Multi-Collector Secondary Ion Mass Spectrometry

2021 ◽  
Vol 8 ◽  
Author(s):  
Nordine Bouden ◽  
Johan Villeneuve ◽  
Yves Marrocchi ◽  
Etienne Deloule ◽  
Evelyn Füri ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Oxygen Isotope ◽  
Spatial Resolution ◽  
Secondary Ion Mass Spectrometry ◽  
Small Radius ◽  
Large Radius ◽  
Ion Mass Spectrometry ◽  
Secondary Ion ◽  
Isotope Measurements

Secondary ion mass spectrometry (SIMS) is a powerful technique for in situ triple oxygen isotope measurements that has been used for more than 30 years. Since pioneering works performed on small-radius ion microprobes in the mid-80s, tremendous progress has been made in terms of analytical precision, spatial resolution and analysis duration. In this respect, the emergence in the mid-90s of the large-radius ion microprobe equipped with a multi-collector system (MC-SIMS) was a game changer. Further developments achieved on CAMECA MC-SIMS since then (e.g., stability of the electronics, enhanced transmission of secondary ions, automatic centering of the secondary ion beam, enhanced control of the magnetic field, 1012Ω resistor for the Faraday cup amplifiers) allow nowadays to routinely measure oxygen isotopic ratios (18O/16O and 17O/16O) in various matrices with a precision (internal error and reproducibility) better than 0.5‰ (2σ), a spatial resolution smaller than 10 µm and in a few minutes per analysis. This paper focuses on the application of the MC-SIMS technique to the in situ monitoring of mass-independent triple oxygen isotope variations.

Direct visualization of a vast cortical calcium compartment in Paramecium by secondary ion mass spectrometry (SIMS) microscopy: possible involvement in exocytosis

1995 ◽  
Vol 108 (5) ◽  
pp. 1895-1909 ◽  
Author(s):  
N. Stelly ◽  
S. Halpern ◽  
G. Nicolas ◽  
P. Fragu ◽  
A. Adoutte
Keyword(s):  
Mass Spectrometry ◽  
Secondary Ion Mass Spectrometry ◽  
Membrane Vesicles ◽  
Cell Periphery ◽  
A Value ◽  
External Reference ◽  
Ion Mass Spectrometry ◽  
First Time ◽  
Secondary Ion

The plasma membrane of ciliates is underlaid by a vast continuous array of membrane vesicles known as cortical alveoli. Previous work had shown that a purified fraction of these vesicles actively pumps calcium, suggesting that alveoli may constitute a calcium-storage compartment. Here we provide direct confirmation of this hypothesis using in situ visualization of total cell calcium on sections of cryofixed and cryosubstituted cells analyzed by SIMS (secondary ion mass spectrometry) microscopy a method never previously applied to protists. A narrow, continuous, Ca-emitting zone located all along the cell periphery was observed on sections including the cortex. In contrast, Na and K were evenly distributed throughout the cell. Various controls confirmed that emission was from the alveoli, in particular, the emitting zone was still seen in mutants totally lacking trichocysts, the large exocytotic organelles docked at the cell surface, indicating that they make no major direct contribution to the emission. Calcium concentration within alveoli was quantified for the first time in SIMS microscopy using an external reference and was found to be in the range of 3 to 5 mM, a value similar to that for sarcoplasmic reticulum. After massive induction of trichocyst discharge, this concentration was found to decrease by about 50%, suggesting that the alveoli are the main source of the calcium involved in exocytosis.

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