A simple approach to single-cell microdissection and molecular analysis.
OBJECTIVE
To develop a simple approach to single-cell microdissection from Papanicolaou-stained smears and to determine its usefulness in molecular diagnosis using archival cytologic material.


METHODS
Culture cells from three cell lines (TT, A549 and Lu65) were used to prepare Papanicolaou-stained smears. Mount-Quick, a mounting medium, was used to establish the method of sorting target single cells from smear slides under direct microscopic observation. Calcitonin receptor gene and codon 12 of the K-ras oncogene were amplified to check the application of single-cell genetic analysis in cytology.


RESULTS
The target single cell could be simply sorted from Papanicolaou-stained smears using the Mount-Quick microdissection technique. Boiling in water was more suitable for preparing DNA from a single cell or from fewer than five cells versus other methods. Analysis of calcitonin receptor gene and K-ras codon 12 demonstrated that molecular analysis was applicable to a single cell or a few cells from archival Papanicolaou-stained smears.


CONCLUSIONS
This approach has significant implications for cytology: (1) it circumvents the limitations of molecular analysis in cytology and makes the combination of molecular analysis and morphologic diagnosis possible in cases with limited materials; (2) the remaining part of the smear is still preserved well for additional analysis; and (3) the approach is simple, economical and practical for cytology as well as histology.
Zhang
Z
Z
Second Department of Pathology, Wakayama Medical College, Japan.
Nakamura
M
M
Taniguchi
E
E
Shan
L
L
Yokoi
T
T
Kakudo
K
K
D004273:DNA, Neoplasm / Q000032:analysis*; D011905:Genes, ras; D006801:Humans; D018003:Receptors, Calcitonin / Q000235:genetics*; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D018003:Receptors, Calcitonin