Developments in in situ hybridisation

Cassidy, Andrew; Jones, Julia

doi:10.1016/j.ymeth.2014.04.006

Cited by 52 publications

(36 citation statements)

References 17 publications

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“…It was replaced by fluorescence-based techniques (FISH) when fluorescent dyes became available (Langer et al, 1981). FISH has been improved significantly in the past decades, providing high efficiency, sensitivity, and spatial resolution (Jiang and Gill, 2006;Volpi and Bridger, 2008;Cassidy and Jones, 2014), and is now used extensively in both biology research and clinical diagnostics.…”

Section: Commentary Background Informationmentioning

confidence: 99%

In Situ Hybridization in Rice (Oryza sativa)

Wang

2016

CP Plant Biology

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Fluorescence in situ hybridization (FISH) is widely used in cytogenetics to determine the localization of DNA sequences on target chromosomes, to provide visible information regarding the physical position of DNA sequences, to determine the abundance and distribution of repetitive sequences that comprise a large proportion of genomes, and to determine the relative chromosome positions of multiple sequences in physical mapping. By mapping on extended chromatin fibers, fiber‐FISH can be used to determine the structure and organization of genes or DNA sequences with a high resolution (to a few kilobases). The protocols described here will provide procedures of FISH on metaphase chromosomes and extended chromatin fibers of rice (Oryza sativa). © 2016 by John Wiley & Sons, Inc.

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Section: Commentary Background Informationmentioning

confidence: 99%

In Situ Hybridization in Rice (Oryza sativa)

Wang

2016

CP Plant Biology

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“…These stains are typically performed on the same automated platforms on which IHC is done. Instead of an antigen-antibody design, ISH entails the use of chromogen-tagged nucleic acids complementary to target DNA or RNA sequences [ 7 ]. Like IHC, ISH permits the identifi cation of target sequences in a tissuespecifi c context.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Laboratory Techniques Used in the Diagnosis of Pediatric Tumors

Hoehn

Loghavi

2014

Pediatric Malignancies: Pathology and Imaging

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“…Its ability to provide a detailed spatial analysis of gene expression and chromosomes at the single cell level, to identify the cell type, and to detect presence of viral genomes in the analyzed sample has led to its extensive use in a wide range of applications. 1,[3][4][5][6][7] FISH reactions are typically performed using a bench-top assay by pipetting the FISH hybridization mix onto the cytological sample, in which the transport of probes into the cell is primarily diffusion-based and hence is slow. 1,8 This leads to typical incubation times of up to 16 hours for low copy number targets 3,9 and 48 to 94 hours for entire genome hybridization.…”

Section: Introductionmentioning

confidence: 99%

“…1,[3][4][5][6][7] FISH reactions are typically performed using a bench-top assay by pipetting the FISH hybridization mix onto the cytological sample, in which the transport of probes into the cell is primarily diffusion-based and hence is slow. 1,8 This leads to typical incubation times of up to 16 hours for low copy number targets 3,9 and 48 to 94 hours for entire genome hybridization. 4 Once hybridization is complete, FISH signals are analyzed using an end-point analysis after a stringent wash of the hybridization buffer containing the fluorescent probes from the surface.…”

Section: Introductionmentioning

confidence: 99%

Real-Time Monitoring of Fluorescence in Situ Hybridization Kinetics

et al. 2018

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We present a novel method for real-time monitoring and kinetic analysis of fluorescence in situ hybridization (FISH). We implement the method using a vertical microfluidic probe containing a microstructure designed for rapid switching between probe solution and nonfluorescent imaging buffer. The FISH signal is monitored in real time during the imaging buffer wash, during which signal associated with unbound probes is removed. We provide a theoretical description of the method as well as a demonstration of its applicability using a model system of centromeric probes (Cen17). We demonstrate the applicability of the method for characterization of FISH kinetics under conditions of varying probe concentration, destabilizing agent (formamide) content, volume exclusion agent (dextran sulfate) content, and ionic strength. We show that our method can be used to investigate the effect of each of these variables and provide insight into processes affecting in situ hybridization, facilitating the design of new assays.

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Developments in in situ hybridisation

Cited by 52 publications

References 17 publications

In Situ Hybridization in Rice (Oryza sativa)

In Situ Hybridization in Rice (Oryza sativa)

Laboratory Techniques Used in the Diagnosis of Pediatric Tumors

Real-Time Monitoring of Fluorescence in Situ Hybridization Kinetics

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