PMID- 9845440
OWN - NLM
STAT- MEDLINE
DCOM- 19990211
LR  - 20191102
IS  - 0196-4763 (Print)
IS  - 0196-4763 (Linking)
VI  - 33
IP  - 4
DP  - 1998 Dec 1
TI  - Use of platinum coproporphyrin and delayed luminescence imaging to extend the 
      number of targets FISH karyotyping.
PG  - 453-9
AB  - Combinatorial use of fluorophores in multicolor fluorescence in situ 
      hybridization (FISH) allows for the recognition of all human chromosomes. Here we 
      introduce the concept of the use of delayed luminescence labels such as 
      phosphorescent platinum coproporphyrins (PtCP) to extend the number of 
      simultaneously detectable targets in multicolor FISH karyotyping. PtCP-conjugated 
      antibodies were used in combination with conventional FISH labels such as cascade 
      blue, fluorescein, lissamine rhodamine, Cy5, and Cy7. Probe sets for all human 
      chromosomes were generated and labeled with these dyes in a combinatorial 
      approach. Delayed luminescence of PtCP was accomplished using a standard 
      fluorescence microscope in which a specially constructed module for visualization 
      of delayed luminescence was incorporated. The module consists of a minichopper 
      incorporated in the standard block that holds the shutter and diaphragm, and a 
      FLC polarizing shutter mounted in a filter holder at the emission side. 
      Multicolor FISH staining was applied to normal metaphase chromosomes and to 
      chromosomes generated from cultured JVM-2 cells with known translocations. 
      Multicolor FISH images (conventional and delayed) were registered using a 
      slow-scan CCD camera. Recognition of all 24 chromosomes was feasible, since the 
      delayed PtCP fluorescence (lifetime, 90 micros) could be easily distinguished 
      from the conventional promptly fluorescing dyes. We discuss possibilities for 
      extending the number of targets far beyond the 24 demonstrated so far.
FAU - Tanke, H J
AU  - Tanke HJ
AD  - Department of Molecular Cell Biology, Leiden University Medical Center, The 
      Netherlands. tanke@rullf2.leidenuniv.nl
FAU - De Haas, R R
AU  - De Haas RR
FAU - Sagner, G
AU  - Sagner G
FAU - Ganser, M
AU  - Ganser M
FAU - van Gijlswijk, R P
AU  - van Gijlswijk RP
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Cytometry
JT  - Cytometry
JID - 8102328
RN  - 0 (Coproporphyrins)
RN  - 49DFR088MY (Platinum)
SB  - IM
MH  - *Coproporphyrins
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Karyotyping
MH  - Luminescent Measurements
MH  - *Platinum
EDAT- 1998/12/09 03:04
MHDA- 2000/06/20 09:00
CRDT- 1998/12/09 03:04
PHST- 1998/12/09 03:04 [pubmed]
PHST- 2000/06/20 09:00 [medline]
PHST- 1998/12/09 03:04 [entrez]
AID - 10.1002/(SICI)1097-0320(19981201)33:4<453::AID-CYTO9>3.0.CO;2-S [pii]
AID - 10.1002/(sici)1097-0320(19981201)33:4<453::aid-cyto9>3.0.co;2-s [doi]
PST - ppublish
SO  - Cytometry. 1998 Dec 1;33(4):453-9. doi: 
      10.1002/(sici)1097-0320(19981201)33:4<453::aid-cyto9>3.0.co;2-s.