Targeting hepatocytes from liver tissue by laser capture microdissection and proteomics expression profiling.

Research output: Contribution to journalArticle

Abstract

A tissue proteomics process is presented where hepatocyte cell isolation in combination with two-dimensional (2-D) gel electrophoresis and mass spectrometric identification were used to annotate the liver proteome. Laser microdissection of 8 m liver tissue sections was performed and protein expression profiling was compared using a variety of quantities of input cells, and gel separation conditions. The 30 m diameter laser generated the highest protein yields from the polymer coated caps following microsolubilization. We found that 6000 laser pulses (approximately 7200 hepatocytes) were required in order to generate high-resolution gel maps. Within homogeneous tissue samples, this could be accomplished in a total cycle time of 20 min using an automated dissection procedure. Close to 1000 high-quality gel annotations were generated from the corresponding 2-D gel expression profiles which matched closely the corresponding patterns of analytical-scale liver preparations detected by silver staining.

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Authors
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Cell and Molecular Biology

Keywords

  • Matrix-assisted laser desorption/ionization-time of flight, Laser microdissection, Liver tissue, Two-dimensional gel electrophoresis, Miniaturization
Original languageEnglish
Pages (from-to)3800-3805
JournalElectrophoresis
Volume24
Issue number21
Publication statusPublished - 2003
Publication categoryResearch
Peer-reviewedYes

Bibliographic note

The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Organic chemistry (S/LTH) (011001240), Biomedical Engineering (011200011), Department of Clinical Sciences, Lund (013230000), Department of Experimental Medical Science (013210000), Department of Chemistry (011001220)