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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/127953
- author
- Marko-Varga, György
LU
; Berglund, Magnus
LU
; Malmström, Johan
LU
; Lindberg, H.
and Fehniger, T E
- organization
- publishing date
- 2003
- 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 Inc.
- external identifiers
-
- wos:000186858700031
- pmid:14613208
- scopus:0345873242
- ISSN
- 0173-0835
- DOI
- 10.1002/elps.200305645
- language
- English
- LU publication?
- yes
- additional info
- 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)
- 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
- 2016-04-01 12:08:05
- date last changed
- 2022-01-26 23:19:35
@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}},
keywords = {{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 Inc.}},
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}},
doi = {{10.1002/elps.200305645}},
volume = {{24}},
year = {{2003}},
}