The Single-Probe: A Miniaturized Multifunctional Device for Single Cell Mass Spectrometry Analysis

Pan, Ning; Rao, Wei; Kothapalli, Naga Rama; Liu, Renmeng; Burgett, Anthony W. G.; Yang, Zhibo

doi:10.1021/ac5029038

Cited by 199 publications

(273 citation statements)

References 39 publications

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“…For SCMS the single-probe was able to achieve the analysis of individual live HeLa cells 16 . The tip size of the single-probe is typically less than 10 μm (Figure 3a), which is small enough to be directly inserted into many types of eukaryotic cells, of which the diameter is ∼10 μm, for extraction and MS analysis.…”

Section: Representative Resultsmentioning

confidence: 99%

“…In addition to the MSI experiments, it is capable of conducting near real-time in-situ SCMS to elucidate detailed chemical information from live eukaryotic cells 16 , which is a major advantage compared with other vacuum based SCMS techniques (such as MALDI 10 and SIMS 21 ). The small size of the probe tip provides the ability to be inserted into a live eukaryotic cell and to extract and ionize the intracellular compounds for immediate MS analysis.…”

Section: Discussionmentioning

confidence: 99%

“…To reduce the chance of device clogging, ensure to avoid touching the nucleus when inserting the single-probe tip into a cell. If clogging occurs, the device can be regenerated by heating up the clogged probe tip or the nano-ESI emitter using a homebuilt heating coil 16 . Another limitation of the single-probe SCMS technique is that only the adhesive cells ( i.e.…”

Section: Discussionmentioning

confidence: 99%

“…d ) A representative list of metabolites detected from SCMS analysis of HeLa cells both in the positive and negative ion modes. e-f ) Mass spectra for the control and treated (100 nM OSW-1) cells (adapted with permission from reference 16). …”

Section: Figurementioning

confidence: 99%

“…The single-probe, a miniaturized multifunctional device that is capable of conducting high spatial resolution ambient MSI on biological tissue sections 15 and near real-time in-situ SCMS on live single cells 16 . The single-probe has an integrated construction that is made up of a pulled dual-bore quartz capillary coupled with a solvent providing inlet and a nano-ESI emitter made from fused silica capillaries, enabling solvent delivery and analyte extraction to be performed from a single device.…”

Section: Introductionmentioning

confidence: 99%

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Applications of the Single-probe: Mass Spectrometry Imaging and Single Cell Analysis under Ambient Conditions

Rao

Pan

Yang

2016

JoVE

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Mass spectrometry imaging (MSI) and in-situ single cell mass spectrometry (SCMS) analysis under ambient conditions are two emerging fields with great potential for the detailed mass spectrometry (MS) analysis of biomolecules from biological samples. The single-probe, a miniaturized device with integrated sampling and ionization capabilities, is capable of performing both ambient MSI and in-situ SCMS analysis. For ambient MSI, the single-probe uses surface micro-extraction to continually conduct MS analysis of the sample, and this technique allows the creation of MS images with high spatial resolution (8.5 μm) from biological samples such as mouse brain and kidney sections. Ambient MSI has the advantage that little to no sample preparation is needed before the analysis, which reduces the amount of potential artifacts present in data acquisition and allows a more representative analysis of the sample to be acquired. For in-situ SCMS, the single-probe tip can be directly inserted into live eukaryotic cells such as HeLa cells, due to the small sampling tip size (< 10 μm), and this technique is capable of detecting a wide range of metabolites inside individual cells at near real-time. SCMS enables a greater sensitivity and accuracy of chemical information to be acquired at the single cell level, which could improve our understanding of biological processes at a more fundamental level than previously possible. The single-probe device can be potentially coupled with a variety of mass spectrometers for broad ranges of MSI and SCMS studies.

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Section: Representative Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Applications of the Single-probe: Mass Spectrometry Imaging and Single Cell Analysis under Ambient Conditions

Rao

Pan

Yang

2016

JoVE

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Single‐Cell Mass Spectrometry

Bergman

Duncan

Lanekoff

2018

Encyclopedia of Analytical Chemistry

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Over the past few decades, the chemical characterization of single cells has improved immensely. In particular, mass spectrometry (MS) has pioneered direct analysis of metabolites, lipids, and peptides from single cells. This progress has been enabled by new and improved strategies for ionization and sampling, where a multitude of techniques for single‐cell MS has contributed unique insights to many different disciplines. Here, an overview of the main three techniques secondary ion mass spectrometry (SIMS), matrix‐assisted laser desorption ionization (MALDI), and ambient ionization for direct single‐cell MS analysis are presented, including some example studies detailing the use of single‐cell MS.

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