Format

Send to

Choose Destination
Nat Commun. 2015 May 12;6:7147. doi: 10.1038/ncomms8147.

Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes.

Author information

1
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
2
1] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA [2] Howard Hughes Medical Institute, Cambridge, Massachusetts 02138, USA.
3
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA [2] Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
4
1] Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA [2] Institut de Génétique Humaine, CNRS UPR 1142, 141 rue de la Cardonille, 34396 Montpellier, France.
5
1] Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA [2] Howard Hughes Medical Institute, Boston, Massachusetts 02114, USA [3] Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
6
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA [2] Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA [3] Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA.
7
1] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA [2] Howard Hughes Medical Institute, Cambridge, Massachusetts 02138, USA [3] Department of Physics, Harvard University, Cambridge, Massachusetts 02138, USA.

Abstract

Fluorescence in situ hybridization (FISH) is a powerful single-cell technique for studying nuclear structure and organization. Here we report two advances in FISH-based imaging. We first describe the in situ visualization of single-copy regions of the genome using two single-molecule super-resolution methodologies. We then introduce a robust and reliable system that harnesses single-nucleotide polymorphisms (SNPs) to visually distinguish the maternal and paternal homologous chromosomes in mammalian and insect systems. Both of these new technologies are enabled by renewable, bioinformatically designed, oligonucleotide-based Oligopaint probes, which we augment with a strategy that uses secondary oligonucleotides (oligos) to produce and enhance fluorescent signals. These advances should substantially expand the capability to query parent-of-origin-specific chromosome positioning and gene expression on a cell-by-cell basis.

PMID:
25962338
PMCID:
PMC4430122
DOI:
10.1038/ncomms8147
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center