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Quantitative mRNA expression analysis in kidney glomeruli using microdissection techniques

Ward De Spiegelaere UGent, Pieter Cornillie UGent, Mario Van Poucke UGent, Luc Peelman UGent, Christian Burvenich UGent and Wim Van Den Broeck UGent (2011) HISTOLOGY AND HISTOPATHOLOGY. 26(2). p.267-275
abstract
The introduction of new tools for molecular analysis, such as RT-qPCR and microarrays, has provided researchers with powerful applications to study renal disease and development. However, the high cellular heterogeneity of the renal tissue complicates the molecular analysis of specific cells and cell groups such as glomerular or tubular cells. In the past, glomerular sieving and manual dissection were used for the isolation of glomeruli. However, these techniques cannot be used for the isolation of specific glomeruli or for the coisolation of additional tissue fractions. In recent decades, new microdissection techniques such as laser-assisted microdissection have been developed. These applications allow the isolation of small cell groups from heterogeneous tissue for molecular analysis, including microarray and RT-qPCR. Although very promising, some drawbacks are associated with these techniques. The isolated sample material is generally small and requires sensitive assays. In addition, the long sample processing time may result in a considerable loss of RNA integrity. Careful optimization and rigorous quality analysis should overcome these drawbacks. In the present paper, the recent literature on the application of microdissection techniques in kidney research is reviewed, together with a discussion of the critical issues that are essential for the application of quantitative mRNA expression analysis with RT-qPCR on microdissected samples.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (review)
publication status
published
subject
keyword
REAL-TIME PCR, LASER-CAPTURE MICRODISSECTION, PARAFFIN-EMBEDDED TISSUES, GENE-EXPRESSION, RT-PCR, Kidney, Glomerular isolation, Gene expression analysis, Real time PCR, Laser microdissection, Glomerular sieving, DEGRADED RNA, EPITHELIAL-CELLS, GROWTH-FACTOR, ASSISTED MICRODISSECTION, MOLECULAR DIAGNOSIS
journal title
HISTOLOGY AND HISTOPATHOLOGY
Histol. Histopath.
volume
26
issue
2
pages
267 - 275
Web of Science type
Article
Web of Science id
000285238800012
JCR category
PATHOLOGY
JCR impact factor
2.48 (2011)
JCR rank
29/78 (2011)
JCR quartile
2 (2011)
ISSN
0213-3911
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1849317
handle
http://hdl.handle.net/1854/LU-1849317
date created
2011-06-30 13:59:57
date last changed
2011-07-27 15:56:08
@article{1849317,
  abstract     = {The introduction of new tools for molecular analysis, such as RT-qPCR and microarrays, has provided researchers with powerful applications to study renal disease and development. However, the high cellular heterogeneity of the renal tissue complicates the molecular analysis of specific cells and cell groups such as glomerular or tubular cells. In the past, glomerular sieving and manual dissection were used for the isolation of glomeruli. However, these techniques cannot be used for the isolation of specific glomeruli or for the coisolation of additional tissue fractions. In recent decades, new microdissection techniques such as laser-assisted microdissection have been developed. These applications allow the isolation of small cell groups from heterogeneous tissue for molecular analysis, including microarray and RT-qPCR. Although very promising, some drawbacks are associated with these techniques. The isolated sample material is generally small and requires sensitive assays. In addition, the long sample processing time may result in a considerable loss of RNA integrity. Careful optimization and rigorous quality analysis should overcome these drawbacks. In the present paper, the recent literature on the application of microdissection techniques in kidney research is reviewed, together with a discussion of the critical issues that are essential for the application of quantitative mRNA expression analysis with RT-qPCR on microdissected samples.},
  author       = {De Spiegelaere, Ward and Cornillie, Pieter and Van Poucke, Mario and Peelman, Luc and Burvenich, Christian and Van Den Broeck, Wim},
  issn         = {0213-3911},
  journal      = {HISTOLOGY AND HISTOPATHOLOGY},
  keyword      = {REAL-TIME PCR,LASER-CAPTURE MICRODISSECTION,PARAFFIN-EMBEDDED TISSUES,GENE-EXPRESSION,RT-PCR,Kidney,Glomerular isolation,Gene expression analysis,Real time PCR,Laser microdissection,Glomerular sieving,DEGRADED RNA,EPITHELIAL-CELLS,GROWTH-FACTOR,ASSISTED MICRODISSECTION,MOLECULAR DIAGNOSIS},
  language     = {eng},
  number       = {2},
  pages        = {267--275},
  title        = {Quantitative mRNA expression analysis in kidney glomeruli using microdissection techniques},
  volume       = {26},
  year         = {2011},
}

Chicago
DE SPIEGELAERE, WARD, Pieter Cornillie, Mario Van Poucke, Luc Peelman, Christian Burvenich, and Wim Van Den Broeck. 2011. “Quantitative mRNA Expression Analysis in Kidney Glomeruli Using Microdissection Techniques.” Histology and Histopathology 26 (2): 267–275.
APA
DE SPIEGELAERE, W., Cornillie, P., Van Poucke, M., Peelman, L., Burvenich, C., & Van Den Broeck, W. (2011). Quantitative mRNA expression analysis in kidney glomeruli using microdissection techniques. HISTOLOGY AND HISTOPATHOLOGY, 26(2), 267–275.
Vancouver
1.
DE SPIEGELAERE W, Cornillie P, Van Poucke M, Peelman L, Burvenich C, Van Den Broeck W. Quantitative mRNA expression analysis in kidney glomeruli using microdissection techniques. HISTOLOGY AND HISTOPATHOLOGY. 2011;26(2):267–75.
MLA
DE SPIEGELAERE, WARD, Pieter Cornillie, Mario Van Poucke, et al. “Quantitative mRNA Expression Analysis in Kidney Glomeruli Using Microdissection Techniques.” HISTOLOGY AND HISTOPATHOLOGY 26.2 (2011): 267–275. Print.