PMID- 9858810
OWN - NLM
STAT- MEDLINE
DCOM- 19990201
LR  - 20180213
IS  - 0301-0171 (Print)
IS  - 0301-0171 (Linking)
VI  - 82
IP  - 3-4
DP  - 1998
TI  - A versatile image analysis approach for simultaneous chromosome identification 
      and localization of FISH probes.
PG  - 172-9
AB  - Modern cytogenetic techniques, such as comparative genomic hybridization (CGH) 
      and the multi-color fluorescence in situ hybridization (FISH) techniques of 
      multiplex fluorescence in situ hybridization (M-FISH) and spectral karyotyping 
      (SKY), require a coordinated banding analysis to maximize their usefulness. All 
      of the methods currently used, including Giemsa (G-) banding, Alu banding, and 
      4',6-diamidino-2-phenyl-indole (DAPI) banding, have serious drawbacks. A simple 
      and effective method to band chromosomes concurrently with FISH is needed. To 
      address this problem, we stained chromosomes with DAPI and chromomycin A3, and 
      then used an image analysis program to generate banding by dividing the image 
      taken with a DAPI excitation filter by the image taken with a chromomycin A3 
      excitation filter. The result was a metaphase spread in which the chromosomes 
      possessed a banding pattern characteristic of R-banding. The image analysis 
      program was then used to generate linescans of pixel intensity versus relative 
      position along the length of chromosomes that were banded using this technique, 
      which we have called D/C R-banding. Each chromosome in a genome was represented 
      by a characteristic scan profile, which was unaffected by FISH signals. Reference 
      linescans were prepared by karyotyping D/C R-banded chromosomes for a given 
      species, and then drawing lines along the length of the known chromosomes. The 
      linescans were combined into a spreadsheet database, which was linked by dynamic 
      data exchange to the image analysis program and normalized for length and 
      intensity. The linescan of an unknown chromosome was then transferred to the 
      spreadsheet, where it was normalized for length and intensity and overlaid on the 
      linescans of each chromosome in the genome. Unknown chromosomes were identified 
      by comparison of their graphs with graphs in the standardized reference genome. 
      We have used this approach to create reference linescan karyotypes of several 
      species, and to identify chromosomes on which FISH was performed.
FAU - Christian, A
AU  - Christian A
AD  - Department of Radiological Health Sciences, Colorado State University, Fort 
      Collins, CO 80523-1673, (USA).
FAU - McNiel, E
AU  - McNiel E
FAU - Robinson, J
AU  - Robinson J
FAU - Drabek, R
AU  - Drabek R
FAU - LaRue, S
AU  - LaRue S
FAU - Waldren, C
AU  - Waldren C
FAU - Bedford, J
AU  - Bedford J
LA  - eng
PT  - Journal Article
PL  - Switzerland
TA  - Cytogenet Cell Genet
JT  - Cytogenetics and cell genetics
JID - 0367735
RN  - 0 (DNA Probes)
SB  - IM
MH  - Animals
MH  - Cats
MH  - DNA Mutational Analysis/methods
MH  - DNA Probes
MH  - Dogs
MH  - Humans
MH  - Image Processing, Computer-Assisted/*methods
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Karyotyping
MH  - Metaphase
OTO - NASA
OT  - Non-programmatic
EDAT- 1998/12/22 03:02
MHDA- 2000/08/16 11:00
CRDT- 1998/12/22 03:02
PHST- 1998/12/22 03:02 [pubmed]
PHST- 2000/08/16 11:00 [medline]
PHST- 1998/12/22 03:02 [entrez]
AID - ccg82172 [pii]
AID - 10.1159/000015093 [doi]
PST - ppublish
SO  - Cytogenet Cell Genet. 1998;82(3-4):172-9. doi: 10.1159/000015093.