Identification of expressed genes by laser-mediated manipulation of single cells

Nat Biotechnol. 1998 Aug;16(8):737-42. doi: 10.1038/nbt0898-737.

Abstract

We describe a rapid noncontact method for the capture of single cells or small tissue areas of any size or shape directly within the cap of a common microfuge tube. Prior to the laser-mediated transfer, the specimen is isolated by laser microbeam microdissection, forming a clear-cut gap around the selected area. Laser treatment does not impair subsequent RNA analysis. We have used this method to isolate a single cell from archival colon adenocarcinoma, and were able to detect point mutations within codon 12 of c-Ki-ras2 mRNA after nested RT-PCR analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / genetics
  • Adenocarcinoma / pathology*
  • Cell Separation / methods*
  • Codon / genetics
  • Colonic Neoplasms / genetics
  • Colonic Neoplasms / pathology*
  • Deoxyribonuclease HpaII
  • Deoxyribonucleases, Type II Site-Specific
  • Genes, ras*
  • Humans
  • Lasers*
  • Microtomy / methods
  • Point Mutation
  • Polymerase Chain Reaction / methods
  • RNA, Messenger / genetics

Substances

  • Codon
  • RNA, Messenger
  • Deoxyribonuclease HpaII
  • Deoxyribonucleases, Type II Site-Specific
  • TCGA-specific type II deoxyribonucleases