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

In situ visualization of proteins of interest at single cell level is attractive in cell biology, molecular biology and biomedicine, which usually involves <a></a><a>photon, electron or X-ray</a> 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 <a>chemical</a> 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 in situ visualization of proteins of interest as well as the interactions between proteins and drugs or drug damaged DNA in single cells.

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

10.26434/chemrxiv.12046125.v2 ◽

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

In situ 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 in situ visualization of proteins of interest as well as the interactions between proteins and drugs or drug damaged DNA in single cells.

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

Analytical Chemistry ◽

10.1021/acs.analchem.0c03448 ◽

2020 ◽

Vol 92 (23) ◽

pp. 15517-15525

Author(s):

Feifei Jia ◽

Jie Wang ◽

Yao Zhao ◽

Yanyan Zhang ◽

Qun Luo ◽

...

Keyword(s):

Mass Spectrometry ◽

Secondary Ion Mass Spectrometry ◽

Single Cells ◽

Time Of Flight ◽

In Situ Visualization ◽

Ion Mass Spectrometry ◽

Chemical Tags ◽

Secondary Ion

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In-situ Detection of Deuterium in Duplex Stainless Steels by Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS)

International Hydrogen Conference (IHC 2012) ◽

10.1115/1.860298_ch54 ◽

2014 ◽

pp. 505-512

Keyword(s):

Mass Spectrometry ◽

Stainless Steels ◽

Secondary Ion Mass Spectrometry ◽

Time Of Flight ◽

Duplex Stainless Steels ◽

Tof Sims ◽

Ion Mass Spectrometry ◽

In Situ Detection ◽

Secondary Ion

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Time-of-flight secondary ion mass spectrometry of bulk polymers using patterned silver overlayers to enhance the molecular microanalysis of trace additives

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010013239x ◽

1992 ◽

Vol 50 (2) ◽

pp. 1552-1553

Author(s):

M. P. Mawn ◽

R. W. Linton ◽

A. Benninghoven

Keyword(s):

Mass Spectrometry ◽

Surface Segregation ◽

Secondary Ion Mass Spectrometry ◽

Time Of Flight ◽

Polymer Additives ◽

Mass Analyzer ◽

Tof Sims ◽

Ion Mass Spectrometry ◽

Secondary Ion

When coupled to a time-of-flight (TOF) mass analyzer which provides a high transmission, quasi-simultaneous detection of secondary ions, and theoretically unlimited mass range, secondary ion mass spectrometry (SIMS) is well suited for submonolayer molecular analysis of solid surfaces. Additionally, molecular ion images may be obtained by using a focused primary ion beam rastered on the sample surface to provide lateral distributions of chemical species.TOF-SIMS has been shown effective at characterizing low level polymer additives such as UV absorbers, thermal stabilizers, anti-oxidants, and slip agents in polymer extracts deposited on roughened Ag substrates. Additionally, in-situ analysis of linear low density polyethylene (LLDPE) samples using TOF-SIMS has provided information on surface segregation and surface oxidation. The polymer extract analysis has the advantage of both preconcentrating the polymer additives, as well as providing enhanced ion yields through Ag canonization. However, the in-situ analysis is also a valuable approach since it alleviates the time consuming extraction procedure and provides information on additive surface segregation and oxidation.

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Applications of Time-OF-Flight Secondary Ion Mass Spectrometry (TOF-SIMS)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100135149 ◽

1990 ◽

Vol 48 (2) ◽

pp. 308-309

Author(s):

Bruno Schueler ◽

Robert W. Odom

Keyword(s):

Mass Spectrometry ◽

Secondary Ion Mass Spectrometry ◽

Structural Information ◽

Time Of Flight ◽

Biological Tissues ◽

Polymer Surfaces ◽

Tof Sims ◽

Secondary Ions ◽

Ion Mass Spectrometry ◽

Secondary Ion

Time-of-flight secondary ion mass spectrometry (TOF-SIMS) provides unique capabilities for elemental and molecular compositional analysis of a wide variety of surfaces. This relatively new technique is finding increasing applications in analyses concerned with determining the chemical composition of various polymer surfaces, identifying the composition of organic and inorganic residues on surfaces and the localization of molecular or structurally significant secondary ions signals from biological tissues. TOF-SIMS analyses are typically performed under low primary ion dose (static SIMS) conditions and hence the secondary ions formed often contain significant structural information.This paper will present an overview of current TOF-SIMS instrumentation with particular emphasis on the stigmatic imaging ion microscope developed in the authors’ laboratory. This discussion will be followed by a presentation of several useful applications of the technique for the characterization of polymer surfaces and biological tissues specimens. Particular attention in these applications will focus on how the analytical problem impacts the performance requirements of the mass spectrometer and vice-versa.

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