Laser microdissection and gene expression analysis on formaldehyde-fixed archival tissue.

Author(s): Cohen CD, Gröne HJ, Gröne EF, Nelson PJ, Schlöndorff D, Kretzler M

Publication: Kidney Int, 2002, Vol. 61, Page 125-32

PubMed ID: 11786092 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine whether mRNA expression levels could be quantified from laser microdissected formaldehyde-fixed or snap-frozen renal tissue using real time RT-PCR.

   Summary of Findings:

   While formaldehyde-fixed tissue sections showed better morphological preservation, laser microdissection and laser pressure catapulting of desired tissue compartments was successful for both formaldehyde-fixed and snap-frozen tissue. Formaldehyde-fixed tissue yielded approximately 5- to 7-fold less mRNA than snap-frozen tissue, however enough mRNA was recovered for real time qRT-PCR by both methods. Further, after normalization to GAPDH, comparable mRNA expression was determined for IP-10 and RANTES.

   Biospecimens

   • Tissue - Kidney

   Preservative Types

   • Frozen
   • Other Preservative

   Diagnoses:

   • Normal
   • Neoplastic - Carcinoma
   • Neoplastic - Normal Adjacent
   • Other diagnoses

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR
   Protein	Immunohistochemistry
   Morphology	Light microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Formaldehyde Snap frozen
   Preaquisition	Diagnosis/ patient condition	Renal transplant rejection Renal carcinoma
   Storage	Storage temperature	Room temperature -80 degrees C
   Biospecimen Aliquots and Components	Cell capture method	Laser microdissection and laser pressure catapulting
   Real-time qRT-PCR Specific	Targeted nucleic acid	IP-10 RANTES WT-1 GAPDH

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