Laser microdissection and pressure catapulting (LMPC) in paraffin sections mounted on glass slides. A methodological report.
The technique of laser microdissection together with laser pressure catapulting (LMPC) is demonstrated in paraffin sections obtained from surgical specimens of brain tumors mounted on glass slides. A sufficient and precise application of microdissection techniques in tissue on glass slides is worthwhile, since it offers the possibility of a retrospective analysis of archived paraffin sections in histopathology. We could demonstrate a precise dissection of areas in tissues of different thicknesses (4 microm and 20 microm). Areas of tissue mounted directly on glass need to be dissected in a scanning mode in order to remove the total region in form of small tissue fragments row by row. This mode provided a precise microdissection of tissue areas of different sizes and shapes. A successful molecular biological analysis of the microdissected regions could be demonstrated. As an example for such an analysis, differential-PCR for detecting an amplification of the gene for the epidermal growth factor receptor (EGFR) was performed.
Nafe
Reinhold
R
Department Neuroradiology, Clinics of Goethe-University, Frankfurt am Main, Germany. r.nafe@em.uni-frankfurt.de
Gangnus
Rainer
R
Glienke
Wolfgang
W
Burgemeister
Renate
R
Haar
Beate
B
Pries
Antje
A
Schlote
Wolfgang
W
D016543:Central Nervous System Neoplasms / Q000473:pathology*; D004210:Dissection / Q000379:methods*; D018773:Genes, erbB-1 / Q000235:genetics; D005910:Glioma / Q000473:pathology*; D006801:Humans; D007834:Lasers; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction