Laser-assisted microdissection applied to frozen surgical pathologic specimens - methodological aspects on RT-PCR.
Laser-assisted microdissection has become a unique technique for an accurate gene-expression profiling analysis in human tissues. The introduction of this approach requires the development of a reliable, efficient, and reproducible procedure for tissue processing. We report a systematic evaluation of the different relevant steps required to obtain sufficient quantity and good quality RNA for reverse transcriptase-polymerase chain reaction when using frozen surgical pathologic tissues as starting material. We propose an optimized and very efficient method with respect to time and effort that can easily be put into practice in research laboratory as well as in any pathology laboratory.
Saal
Isabelle
I
Laboratory of Pathology, Erasmus University Hospital, Université Libre de Bruxelles, Brussels, Belgium.
Gustin
Anne
A
Rombaut
Katia
K
Pelaez
Pilar
P
Rorive
Sandrine
S
Remmelink
Myriam
M
Salmon
Isabelle
I
D003106:Colon / Q000473:pathology; D005629:Frozen Sections / Q000592:standards*; D006801:Humans; D053685:Laser Therapy / Q000379:methods; D008168:Lung / Q000473:pathology; D042282:Microdissection / Q000379:methods*; D009369:Neoplasms / Q000235:genetics* / Q000473:pathology; D012334:RNA, Neoplasm / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards*; D013961:Thyroid Gland / Q000473:pathology
D012334:RNA, Neoplasm