title: Towards many colors in FISH on 3D-preserved interphase nuclei.
abstract: The article reviews the existing methods of multicolor FISH on nuclear targets, first of all, interphase chromosomes. FISH proper and image acquisition are considered as two related components of a single process. We discuss (1) M-FISH (combinatorial labeling + deconvolution + wide-field microscopy); (2) multicolor labeling + SIM (structured illumination microscopy); (3) the standard approach to multicolor FISH + CLSM (confocal laser scanning microscopy; one fluorochrome - one color channel); (4) combinatorial labeling + CLSM; (5) non-combinatorial labeling + CLSM + linear unmixing. Two related issues, deconvolution of images acquired with CLSM and correction of data for chromatic Z-shift, are also discussed. All methods are illustrated with practical examples. Finally, several rules of thumb helping to choose an optimal labeling + microscopy combination for the planned experiment are suggested.
authors.affiliation: Chair of Anthropology and Human Genetics, Department of Biology II, Ludwig-Maximilians-University, Martinsried, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Walter
authors.forename: B
authors.initials: B
authors.lastname: Joffe
authors.forename: A
authors.initials: A
authors.lastname: Bolzer
authors.forename: H
authors.initials: H
authors.lastname: Albiez
authors.forename: P A
authors.initials: PA
authors.lastname: Benedetti
authors.forename: S
authors.initials: S
authors.lastname: Müller
authors.forename: M R
authors.initials: MR
authors.lastname: Speicher
authors.forename: T
authors.initials: T
authors.lastname: Cremer
authors.forename: M
authors.initials: M
authors.lastname: Cremer
authors.forename: I
authors.initials: I
authors.lastname: Solovei
mesh_terms: D002467:Cell Nucleus / Q000235:genetics / Q000648:ultrastructure*; D020223:Chromosome Painting; D003116:Color; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007399:Interphase / Q000235:genetics*; D008856:Microscopy, Fluorescence