Laser-assisted preparation of single cells from stained histological slides for gene analysis.
Individual cells are prepared from histological tissue sections of routinely formalin-fixed and paraffin-embedded tissues using an ultraviolet laser micromanipulator. This technology, in combination with polymerase chain reaction (PCR)-based gene analysis, will enable researchers to routinely detect a variety of nucleic acid abnormalities underlying cancer, infection, and genetic disease with previously unknown sensitivity: at the single cell level. The utility of this technique is demonstrated by PCR amplification and sequencing of the E-cadherin gene, which codes for a homophilic cell-to-cell adhesion molecule, in early gastric carcinomas of the diffuse type of Lauren's classification. The main characteristics of the laser-assisted microdissection technique are high precision without contamination and easy application. The assignment of individual gene sequences to single cells will now provide a direct link between molecular biology on the one hand and histology and pathology on the other.
Becker
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I
Technische Universität München, Klinikum rechts der Isar, Institut für Pathologie, Germany.
Becker
K F
KF
Röhrl
M H
MH
Höfler
H
H
D015820:Cadherins / Q000235:genetics*; D002469:Cell Separation / Q000379:methods; D003582:Cytogenetics / Q000379:methods*; D004273:DNA, Neoplasm; D005240:Feasibility Studies; D005784:Gene Amplification; D006652:Histological Techniques; D006801:Humans; D007834:Lasers; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013274:Stomach Neoplasms / Q000235:genetics / Q000473:pathology
D015820:Cadherins; D004273:DNA, Neoplasm