Enabling high-sensitivity live single-cell mass spectrometry using an integrated electrical lysis and nano electrospray ionization interface
Analytica Chimica Acta, vol. 1324, pp. 343068
Abstract
BACKGROUND: Live single-cell metabolomic studies encounter inherent difficulties attributed to the limited sample volume, minimal compound quantity, and insufficient sensitivity in the Mass Spectrometry (MS) method used to obtain single-cell data. However, understanding cellular heterogeneity, functional diversity, and metabolic processes within individual cells is essential. Exploring how individual cells respond to stimuli, including drugs, environmental changes, or signaling molecules, offers insights into biology, oncology, and drug discovery. Efficient release of cell contents (lysis) is vital for accurate metabolite detection at the single-cell level. Despite this, traditional approaches in live single cell metabolomics methods do not emphasize efficient lysis to prevent sample dilution. Instead, current live single cell metabolomics methods use direct infusion to introduce the cell into the mass spectrometry without prior chromatographic separation or a lysis step, which adversely affects sensitivity and metabolic coverage. RESULTS: To address this, we developed an integrated single-cell electrical lysis and nano spray (SCEL-nS) platform coupled to an Orbitrap MS capable of efficiently lysing a single cell after being sampled with specially manufactured micropipettes. Lysis efficiency was validated by comparing live cell stain fluorescent intensities of intact and electrically lysed cells through microscopy imaging. The SCEL-nS platform successfully induced the breakdown of a single cell, significantly reducing the live cell stain's fluorescent intensity indicating cell membrane breakdown. Additionally, SCEL-nS was validated by measuring single cells spiked with the anti-cancer drug tamoxifen by MS. SCEL-nS use resulted in statistically significant increase in the peak measured by the method compared to the traditional non-lysis method. SIGNIFICANCE: Overall, our results demonstrate that the newly incorporated SCEL-nS platform achieved higher sensitivities compared to traditional live single cell analysis methods.
Authors 9
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Affiliation as printed
Metabolomics and Analytics Center, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands
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Luís Daniel de Aguiar Homem e Almeida de Matos Aachen Metabolomics and Analytics Centre Leiden Academic Centre for Drug Research
Affiliation as printed
Metabolomics and Analytics Center, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands
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Affiliation as printed
Metabolomics and Analytics Center, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands
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Affiliation as printed
Fine Mechanical Department, Leiden Institute of Physics (LION), Leiden University, Leiden, the Netherlands
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Affiliation as printed
Metabolomics and Analytics Center, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands
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Birla Institute of Technology and Science, Pilani
Affiliation as printed
Department of Mechanical Engineering, Birla Institute of Technology and Science, Pilani, Hyderabad, India
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Affiliation as printed
Metabolomics and Analytics Center, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands
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Ahmed Ali corresponding Aachen Metabolomics and Analytics Centre Leiden Academic Centre for Drug Research
Affiliation as printed
Metabolomics and Analytics Center, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands. Electronic address: ali@lacdr.leidenuniv.nl
Metabolomics and Analytics Center, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands
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Thomas Hankemeier corresponding Aachen Metabolomics and Analytics Centre Leiden Academic Centre for Drug Research
Affiliation as printed
Metabolomics and Analytics Center, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands
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