[Flow cytometric quantification of numerical chromosome aberrations in non-small cell lung carcinomas using formalin-fixed paraffin-embedded tissue].
Fluorescence in situ hybridization with biotinylated repetitive DNA probe specific for the centromeric region of chromosome 17 (p17H8: Oncor) was applied to suspended nuclei which were isolated by Shutte's method from formalin-fixed paraffin-embedded tissue. The tissues were obtained from surgically resected specimens from nine patients with non-small cell lung carcinoma. The isolated nuclei were prepared with 0.05% pepsin/0.1NHCl for 15 minutes at 37 degrees C. Subsequently, these were immersed in 70% acetic acid for 10 seconds at room temperature. After heat denature with hybridization mixture which contained 3 mu 1 DNA probe for 10 minutesat 70 degrees C, 1 x 10(6) nuclei were incubated overnight at 37 degrees C. After washing with 60% formamide/2 x SSC, the hybridized probes were labeled by FITC conjugated avidin. A number of centromeric signals of chromosome 17 wasevaluated by fluorescence microscopy (BH-2, Olympus). Furthermore, a probe-related FITC intensity was quantified using flow cytometry (FACScan, Becton Dickinson). As the results, there was good correlation between a relative fluorescence intensity determined by flow cytometry and a relative fluorescence signal by fluorescence microscopy (p < 0.05).
Sawai
T
T
First Department of Surgery, Nagasaki University School of Medicine, Japan.
Sasano
O
O
Shibazaki
S
S
Shibata
Y
Y
Tsuji
T
T
Nanashima
A
A
Yamaguchi
H
H
Yasutake
T
T
Hara
S
S
Tagawa
Y
Y
Nakagoe
T
T
Ayabe
H
H
D000368:Aged; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D002869:Chromosome Aberrations; D002886:Chromosomes, Human, Pair 17; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008175:Lung Neoplasms / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D016612:Paraffin Embedding / Q000379:methods; D011003:Ploidies
D004273:DNA, Neoplasm