Using Dicationic Ion-Pairing Compounds To Enhance the Single Cell Mass Spectrometry Analysis Using the Single-Probe:
Ning Pan1, Wei Rao1, Shawna J Standke1
1Department of Chemistry and Biochemistry, University of Oklahoma , Norman, Oklahoma 73019, United States.
This study introduces a novel mass spectrometry (MS) method using dicationic ion-pairing compounds to detect negatively charged metabolites in single cells. This technique enhances sensitivity and allows for detailed analysis of cellular components in real-time.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Cell Biology
Background:
- Detecting negatively charged species in live cells using mass spectrometry (MS) presents significant challenges.
- Existing methods often lack the sensitivity or specificity required for single-cell analysis.
- Real-time monitoring of cellular metabolites is crucial for understanding cellular processes.
Purpose of the Study:
- To develop and validate a novel MS method for sensitive detection of negatively charged metabolites in live single cells.
- To utilize dicationic ion-pairing compounds for enhanced signal detection in positive ionization mode.
- To demonstrate the capability of the single-probe MS technique for comprehensive metabolite profiling at the single-cell level.
Main Methods:
- A miniaturized, multifunctional single-probe device was employed for reactive MS analysis.
- Two dicationic reagents, C5(bpyr)2F2 and C3(triprp)2F2, were used as ion-pairing agents.
- Live single cells were analyzed under ambient conditions using positive ionization mode MS.
Main Results:
- The method successfully detected negatively charged metabolites as stable ion-pairs with dicationic compounds in positive ion mode.
- A significant increase in sensitivity was achieved for the detection of cellular metabolites.
- Tentative assignments of 285 metabolites using C5(bpyr)2F2 and 143 using C3(triprp)2F2 were made.
- Selectivity of dicationic compounds enabled discrimination of overlapped ion peaks.
- Tandem MS/MS analyses confirmed molecular identification of selected adduct ions.
Conclusions:
- The developed dicationic ion-pairing MS method provides an effective approach for detecting a wide range of metabolites in single cells.
- This technique significantly improves sensitivity and selectivity for analyzing negatively charged species.
- The single-probe MS platform offers a powerful tool for real-time, in-depth cellular analysis.
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