PMID- 15696183
OWN - NLM
STAT- MEDLINE
DCOM- 20050526
LR  - 20230216
IS  - 0023-6837 (Print)
IS  - 0023-6837 (Linking)
VI  - 85
IP  - 4
DP  - 2005 Apr
TI  - Sequential application of interphase-FISH and CGH to single cells.
PG  - 582-92
AB  - A comprehensive genomic analysis of single cells is needed for numerous scenarios 
      in tumor genetics, clinical diagnostics and forensic application. PCR protocols 
      were developed which allow an unbiased amplification of the whole genome of a 
      single cell for subsequent analyses by comparative genomic hybridization (CGH). 
      However, verification of single-cell CGH results has been impossible as the 
      procedure naturally involves the destruction of the respective cell. Here we show 
      that the genome of individual cells can be analyzed by two different single cell 
      techniques applied sequentially to the same cell. In a first step, interphase 
      fluorescence in situ hybridization (FISH) is applied. After evaluation of the 
      interphase-FISH signals, cells of interest can be selected for a further 
      analysis. Single cells are collected by laser microdissection, the DNA is 
      amplified by linker-adaptor PCR and subjected to CGH-analysis. This strategy 
      offers new opportunities for a sophisticated selection of cells based on 
      interphase-FISH signals. Furthermore, the sequential application of two different 
      single-cell approaches to the same single-cell represents the only option to 
      control and verify the single-cell CGH results. We demonstrate the feasibility of 
      this approach with a series of experiments including cells from pre- and 
      postnatal diagnostics, for example, cells with trisomies 13, 18, or 21, 
      respectively, leukemia and tumor cells and tissue sections.
FAU - Langer, Sabine
AU  - Langer S
AD  - Institut fur Humangenetik, Technische Universitat Munchen, Munich, Germany.
FAU - Geigl, Jochen B
AU  - Geigl JB
FAU - Gangnus, Rainer
AU  - Gangnus R
FAU - Speicher, Michael R
AU  - Speicher MR
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Lab Invest
JT  - Laboratory investigation; a journal of technical methods and pathology
JID - 0376617
SB  - IM
MH  - *In Situ Hybridization, Fluorescence
MH  - *Interphase
MH  - *Nucleic Acid Hybridization
EDAT- 2005/02/08 09:00
MHDA- 2005/05/27 09:00
CRDT- 2005/02/08 09:00
PHST- 2005/02/08 09:00 [pubmed]
PHST- 2005/05/27 09:00 [medline]
PHST- 2005/02/08 09:00 [entrez]
AID - S0023-6837(22)60122-3 [pii]
AID - 10.1038/labinvest.3700249 [doi]
PST - ppublish
SO  - Lab Invest. 2005 Apr;85(4):582-92. doi: 10.1038/labinvest.3700249.