This third edition provides protocols on fluorescence in situ hybridization (FISH) and comparative genomic hybridization approaches. Chapters provide new and updated methods on probe, cell types, tissues, organisms, plants, microorganisms, multicolor FISH procedures, chromosomes, breakpoints, cryptic aberrations, nuclear architecture, epigenetic changes, and comparative genomic hybridization studies, immunostaining, single cell electrophoresis, microdissection, sequencing, and CRISP/Cas9 technology.

Authoritative and cutting-edge, Fluorescence In Situ Hybridization (FISH): Application Guide, Thid Edition aims to be useful to ** researchers just starting to study FISHers and for FISH experts.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Background.- Microscopy and imaging: Visualization matters.- Optical filters and light sources for FISH.- Automation of FISH-procedure and evaluation.- Classification of FISH probes.- Commercial FISH probes.- Generation of painting probes from flow-sorted and microdissected chromosomes.- FISH-microdissection.- Homemade locus-specific FISH probes: Bacterial artificial chromosomes.- The standard FISH procedure.- Microwave treatment for better FISH results in a shorter time.- FISH with and without COT DNA.- One-day FISH procedure.- Quantitative FISH (Q-FISH).- Telomere length measurement by FISH: Image processing techniques.- Directional Genomic Hybridization (dGH) for high-resolution detection of intrachromosomal rearrangements.- The replicative detargeting FISH (ReD-FISH) technique in studies of telomere replication.- Pre- and postnatal diagnostics and research on peripheral blood, bone marrow chorion, amniocytes, and fibroblasts.- Application of FISH to previously GTG-banded and/or embedded cytogenetic slides.- FISH in native amniocytes.- Tumorcytogenetic diagnostics and research on blood and bone marrow smears or effusions.- Characterization of mosaicism in different easy-to-acquire body tissues such as buccal smears, skin abrasions, hair root cells or urine.- Characterization of archived formalin-fixed/paraffin-embedded or cryofixed tissue, including nucleus extraction.- FISH on sperm, spermatocytes and oocytes.- Fluorescence in situ hybridization on DNA fibers: Molecular combing.- Two- to three-color FISH.- Multiplex FISH and spectral karyotyping .- FISH banding techniques.- Centromere directed M-FISH approaches (cenM-FISH).- Heterochromatin directed M-FISH (HCM-FISH).- FISH for repeatedly observed euchromatic variants.- Subtelomeric and/or subcentromeric probe sets.- Bar-coding is more actual than ever.- FISH for repeatedly observed constitutional rearrangements.- Parental origin determination FISH: Pod-FISH.- Simultaneous FISH and fluorescence immunostaining for DNAprotein co-detection.- CENP-antibodies used additionally to FISH.- X-inactivation accessed by FISH and EDU-immunostaining.- Immuno-FISH and microspreading technique for synaptonemal complex studies.- Yeast chromosomes revealed by immuno FISH.- Interphase FISH in diagnostics.- Interphase FISH for detection of chromosomal mosaicism and instability.- Comet-FISH.- Micronucleus FISH.- Three-dimensional interphase analysis using suspension FISH.- 3D DNA-FISH protocol for the study of the DUX gene in preimplantation bovine embryos.- Animal probes and Zoo-FISH.- Three-dimensional co-immuno and RNA-fluorescence in situ hybridization in preimplantation mouse embryos.- FISH in bird chromosomes: An updated protocol and best practices.- An updated standard protocol for FISH mapping in fish species (fish-FISH).- FISH in lampbrush-chromosomes.- General protocol for FISH on insect chromosomes.- FISH in Drosophila.- FISH on insect cells transfected with heterologous DNA a single cell directed approach.- Fluorescence in situ hybridization for plants: Methods of chromosomal preparation.- Mitochondrial DNA (MtDNA) detection by FISH.- Advances in the application of FISH in food microbiology.- Evaluation of polymicrobial involvement using Fluorescence In Situ Hybridization (FISH) in clinical practice.- Comparative genomic hybridization (CGH) and microdissection-based CGH (micro-CGH) in human.- Comparative genomic hybridization (CGH) in animals.- Breakpoint mapping of balanced chromosomal rearrangements using array-CGH with microdissection-derived FISH probes.- Array CGH.- Chromosome microarray results to be verified by FISH.- Sequencing of microdissection derived FISH-probes.- Next generation sequencing should consider molecular cytogenetic results.- Optical genome mapping results need to be verified by FISH.- Application of ALFA-tag signal amplification to expand the CRISPR-based DNA imaging toolkit.