PMID- 22331009
OWN - NLM
STAT- MEDLINE
DCOM- 20120521
LR  - 20211021
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 60
DP  - 2012 Feb 6
TI  - Chromosomics: detection of numerical and structural alterations in all 24 human 
      chromosomes simultaneously using a novel OctoChrome FISH assay.
LID - 3619 [pii]
LID - 10.3791/3619 [doi]
AB  - Fluorescence in situ hybridization (FISH) is a technique that allows specific DNA 
      sequences to be detected on metaphase or interphase chromosomes in cell 
      nuclei(1). The technique uses DNA probes with unique sequences that hybridize to 
      whole chromosomes or specific chromosomal regions, and serves as a powerful 
      adjunct to classic cytogenetics. For instance, many earlier studies reported the 
      frequent detection of increased chromosome aberrations in leukemia patients 
      related with benzene exposure, benzene-poisoning patients, and healthy workers 
      exposed to benzene, using classic cytogenetic analysis(2). Using FISH, 
      leukemia-specific chromosomal alterations have been observed to be elevated in 
      apparently healthy workers exposed to benzene(3-6), indicating the critical roles 
      of cytogentic changes in benzene-induced leukemogenesis. Generally, a single FISH 
      assay examines only one or a few whole chromosomes or specific loci per slide, so 
      multiple hybridizations need to be conducted on multiple slides to cover all of 
      the human chromosomes. Spectral karyotyping (SKY) allows visualization of the 
      whole genome simultaneously, but the requirement for special software and 
      equipment limits its application(7). Here, we describe a novel FISH assay, 
      OctoChrome-FISH, which can be applied for Chromosomics, which we define here as 
      the simultaneous analysis of all 24 human chromosomes on one slide in human 
      studies, such as chromosome-wide aneuploidy study (CWAS)(8). The basis of the 
      method, marketed by Cytocell as the Chromoprobe Multiprobe System, is an 
      OctoChrome device that is divided into 8 squares, each of which carries three 
      different whole chromosome painting probes (Figure 1). Each of the three probes 
      is directly labeled with a different colored fluorophore, green (FITC), red 
      (Texas Red), and blue (Coumarin). The arrangement of chromosome combinations on 
      the OctoChrome device has been designed to facilitate the identification of the 
      non-random structural chromosome alterations (translocations) found in the most 
      common leukemias and lymphomas, for instance t(9;22), t(15;17), t(8;21), 
      t(14;18)(9). Moreover, numerical changes (aneuploidy) in chromosomes can be 
      detected concurrently. The corresponding template slide is also divided into 8 
      squares onto which metaphase spreads are bound (Figure 2), and is positioned over 
      the OctoChrome device. The probes and target DNA are denatured at 
      high-temperature and hybridized in a humid chamber, and then all 24 human 
      chromosomes can be visualized simultaneously. OctoChrome FISH is a promising 
      technique for the clinical diagnosis of leukemia and lymphoma and for detection 
      of aneuploidies in all chromosomes. We have applied this new Chromosomic approach 
      in a CWAS study of benzene-exposed Chinese workers(8,10).
FAU - Ji, Zhiying
AU  - Ji Z
AD  - Genes and Environment Laboratory, University of California, Berkeley, USA.
FAU - Zhang, Luoping
AU  - Zhang L
LA  - eng
GR  - R01 ES017452/ES/NIEHS NIH HHS/United States
GR  - R01ES017452/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Video-Audio Media
DEP - 20120206
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Aneuploidy
MH  - *Chromosome Aberrations
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Leukemia/genetics
MH  - Lymphoma/genetics
PMC - PMC3369631
EDAT- 2012/02/15 06:00
MHDA- 2012/05/23 06:00
PMCR- 2014/02/06
CRDT- 2012/02/15 06:00
PHST- 2012/02/15 06:00 [entrez]
PHST- 2012/02/15 06:00 [pubmed]
PHST- 2012/05/23 06:00 [medline]
PHST- 2014/02/06 00:00 [pmc-release]
AID - 3619 [pii]
AID - 10.3791/3619 [doi]
PST - epublish
SO  - J Vis Exp. 2012 Feb 6;(60):3619. doi: 10.3791/3619.