A microdissection technique for archival DNA analysis of specific cell populations in lesions < 1 mm in size.
We have developed a microdissection technique that allows for procurement and analysis of specific, minute cell populations from routine, 5-mu, formalin-fixed, paraffin-embedded histological tissue sections. Lesions < 1 mm in size can be specifically examined. Cells of interest are procured under direct microscopic visualization followed by a single-step DNA extraction and subsequent polymerase chain reaction. Amplification of DNA from selected cell populations was demonstrated by detecting a loss of heterozygosity (LOH) at the von Hippel-Lindau disease (VHL) gene in an atypical renal lesion and a renal cell carcinoma in a kidney of a VHL patient. Moreover, previously unrecognized LOH on the short arm of chromosome 3 (3p25-26) was detected in microdissected colorectal carcinoma cells in a non-VHL patient with sporadic colon carcinoma. This technique should prove useful in DNA studies of small lesions and cell populations. Furthermore, microscopic premalignant, in situ, and invasive lesions can be selectively examined.
Zhuang
Z
Z
Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Bertheau
P
P
Emmert-Buck
M R
MR
Liotta
L A
LA
Gnarra
J
J
Linehan
W M
WM
Lubensky
I A
IA
D001483:Base Sequence; D002292:Carcinoma, Renal Cell / Q000235:genetics / Q000473:pathology; D002893:Chromosomes, Human, Pair 3 / Q000235:genetics; D003110:Colonic Neoplasms / Q000235:genetics / Q000473:pathology; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D004210:Dissection / Q000379:methods*; D005796:Genes; D006579:Heterozygote; D006801:Humans; D007668:Kidney / Q000473:pathology; D007680:Kidney Neoplasms / Q000235:genetics / Q000473:pathology; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D006623:von Hippel-Lindau Disease / Q000235:genetics
D015335:Molecular Probes; D004247:DNA