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

Marko-Varga, György LU ; Berglund, Magnus LU ; Malmström, Johan LU ; Lindberg, H. and Fehniger, T E (2003) In Electrophoresis 24(21). p.3800-3805
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Matrix-assisted laser desorption/ionization-time of flight, Laser microdissection, Liver tissue, Two-dimensional gel electrophoresis, Miniaturization
in
Electrophoresis
volume
24
issue
21
pages
3800 - 3805
publisher
John Wiley & Sons
external identifiers
  • wos:000186858700031
  • pmid:14613208
  • scopus:0345873242
ISSN
0173-0835
DOI
10.1002/elps.200305645
language
English
LU publication?
yes
id
c9545ea4-efd1-4c18-9e28-eeb6b23d435c (old id 127953)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=14613208&dopt=Abstract
date added to LUP
2007-07-10 13:05:12
date last changed
2018-01-07 05:49:53
@article{c9545ea4-efd1-4c18-9e28-eeb6b23d435c,
  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.},
  author       = {Marko-Varga, György and Berglund, Magnus and Malmström, Johan and Lindberg, H. and Fehniger, T E},
  issn         = {0173-0835},
  keyword      = {Matrix-assisted laser desorption/ionization-time of flight,Laser microdissection,Liver tissue,Two-dimensional gel electrophoresis,Miniaturization},
  language     = {eng},
  number       = {21},
  pages        = {3800--3805},
  publisher    = {John Wiley & Sons},
  series       = {Electrophoresis},
  title        = {Targeting hepatocytes from liver tissue by laser capture microdissection and proteomics expression profiling.},
  url          = {http://dx.doi.org/10.1002/elps.200305645},
  volume       = {24},
  year         = {2003},
}