Automated cell-by-cell tissue imaging and single-cell analysis for targeted morphologies by laser ablation electrospray ionization mass spectrometry

Hang Li, Brian K. Smith, Bindesh Shrestha, L. Márk, Akos Vertes

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Citations (Scopus)

Abstract

Mass spectrometry imaging (MSI) is an emerging technology for the mapping of molecular distributions in tissues. In most of the existing studies, imaging is performed by sampling on a predefined rectangular grid that does not reflect the natural cellular pattern of the tissue. Delivering laser pulses by a sharpened optical fiber in laser ablation electrospray ionization (LAESI) mass spectrometry (MS) has enabled the direct analysis of single cells and subcellular compartments. Cell-by-cell imaging had been demonstrated using LAESI-MS, where individual cells were manually selected to serve as natural pixels for tissue imaging. Here we describe a protocol for a novel cell-by-cell LAESI imaging approach that automates cell recognition and addressing for systematic ablation of individual cells. Cell types with particular morphologies can also be selected for analysis. First, the cells are recognized as objects in a microscope image. The coordinates of their centroids are used by a stage-control program to sequentially position the cells under the optical fiber tip for laser ablation. This approach increases the image acquisition efficiency and stability, and enables the investigation of extended or selected tissue areas. In the LAESI process, the ablation events result in mass spectra that represent the metabolite levels in the ablated cells. Peak intensities of selected ions are used to represent the metabolite distributions in the tissue with single-cell resolution.

Original languageEnglish
Title of host publicationMass Spectrometry Imaging of Small Molecules
PublisherSpringer Fachmedien
Pages117-127
Number of pages11
ISBN (Print)9781493913572, 9781493913565
DOIs
Publication statusPublished - Oct 31 2014

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Keywords

  • Cell-by-cell imaging
  • Imaging
  • Mass spectrometry
  • Metabolites
  • Molecular imaging
  • Single-cell analysis
  • Tissue imaging

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Chemistry(all)

Cite this

Li, H., Smith, B. K., Shrestha, B., Márk, L., & Vertes, A. (2014). Automated cell-by-cell tissue imaging and single-cell analysis for targeted morphologies by laser ablation electrospray ionization mass spectrometry. In Mass Spectrometry Imaging of Small Molecules (pp. 117-127). Springer Fachmedien. https://doi.org/10.1007/978-1-4939-1357-2_12