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, László Márk, Akos Vertes

Research output: Contribution to journalArticle

6 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 predefi ned rectangular grid that does not refl ect the natural cellular pattern of the tissue. Delivering laser pulses by a sharpened optical fi ber 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 fi ber tip for laser ablation. This approach increases the image acquisition effi ciency 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
Pages (from-to)117-127
Number of pages11
JournalMethods in Molecular Biology
Volume1203
DOIs
Publication statusPublished - Jan 1 2015

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Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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