New York – Apr 2nd, 2018 – Fluorescence in situ hybridization, which is a selective assay for the localization of a specific nucleic acid sequence in its natural context. It is a technology that has been in existence for more than 20 years and continues to evolve. During its maturation, various methods and modifications have been introduced to optimize the detection of DNA and RNA. The reasons that make this technology become prevalence are the wide range of applications and its relative ease of implementation and in-situ performance.
Although the basic principles of FISH remain stable, high sensitivity detection, simultaneous determination of multiple species, and automated data collection and analysis have significantly improved this technology. At present, the introduction of FISH surpasses the previously available technologies and becomes the most important biometric. In the future, this technology may have a significant impact on live cell imaging and medical diagnostics.
The Past of fluorescence in situ hybridization
As early as the 1940s, antibodies were conjugated to fluorescent dyes without losing their epitope binding specificity. Several decades later, primary antibody-dependent fluorescence detection of nucleic acid hybrids was achieved, however, this technique was soon replaced by the appearance of fluorescent nucleic acid probes.In situ hybridization, first performed in the late 1960s, was not fluorescent at all, but used a probe labeled with a radioactive isotope. Non-fluorescent technologies such as enzyme-based color reporters and gold-based probe systems used in electron microscopy are all independent.
FISH for visualizing nucleic acids was developed as an alternative method using radiolabeled probes.Early isotope detection methods used non-specific labeling strategies such as random incorporation of radioactively modified bases into growing cells followed by autoradiography. Several disadvantages of isotope hybridization, such as unstable, limited resolution, time-consuming, expensive and difficult to store, have inspired the development of new technologies. The first application of fluorescence in-situ detection was in the early 1980s, detection of nick translations characterized by secondary detection of biotinylated probes and fluorescent streptavidin conjugates was used to detect DNA and mRNA targets. About a decade later, improved labeling of synthetic single-stranded DNA probes allows the chemical preparation of hybridization probes carrying enough fluorescent molecules to allow direct detection. Since then, many changes in the topics of indirect and direct labels have been introduced, providing a wide range of detection solutions for the user to choose.
Although the number of detection methods has increased, the types of targets also have become quite diverse. FISH chromosome analysis has promoted significant advances in cytogenetic studies. However, due to this assay cannot benefit from the preservation of tissue structures or cell structures, its future applications are more likely to be in silico than in situ. Initially, RNA assays can reliably detect considerable amounts of information with clone-derived probes.
The new goal has led to the new application of the FISH program, the popularity of which has increased dramatically in the 1990s. The new research approaches opened up by these applications require the simultaneous detection of more and more species. Initially, this was achieved by simultaneously observing fluorophores of different spectra. Later, strategies that using two major coding schemes expanded the approach.
Prospects for future development
The development of in situ techniques provides us with valuable information about the location and expression patterns of genes in individual cells. A complete gene expression profile of single cells will provide a deeper understanding of the correlation between gene expression patterns and specific cell phenotypes, which is especially important in studies of development and disease progression, where complex, finely-divided gene expression programs are playing a role.
Researchers can predict how molecular pathology ultimately exceeds the limitations of morphopathology, which will allow the more judicious use of minimally invasive biopsy techniques that sacrifice retrieved tissue morphology in favor of comfort of the patient. FISH probes have already colored the way that we visualize and conceptualize genes, chromosomes, transcription and nucleic acid movements. What remains to be seen is how detailed molecular analysis of single cell and tissue samples will influence how we identify, diagnose, and alter the genetic pathology.
In the long run, it is expected that the database of associated gene expression patterns will accumulate at the single cell level as researchers and industry adopt FISH technology and their favorite biomarkers. Ultimately, FISH will be the preferred method for predicting the complex components of genes that cause disease.
About Fluorescence in situ hybridization offered by Creative Bioarray
Creative Bioarray is a leading company in histology service. We are dedicated to providing the most comprehensive list of histology service and custom design services to academia as well as industrial researchers and clinical institutes all around the world. We offer Fluorescence in situ hybridization (FISH) service range from standardized testing of validated assays to custom development of new assays. Drawing on many years of experiences and in-depth knowledge, we guarantee the speed, quality, and cost of our service. Creative Bioarray is your first and most reliable choice in histology.
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