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Fish Technique: Kirti Shikha

FISH is a cytogenetic technique that uses fluorescent probes that bind to chromosomes to locate specific genes or portions of DNA. It allows researchers to visualize genetic material in individual cells. A probe is made complementary to the target sequence and labeled with a fluorescent marker. The chromosomes are treated so the probe can hybridize to its matching sequence. Under a fluorescent microscope, the probe appears as fluorescent signals, showing where the target sequence is located on the chromosomes. FISH has many applications in research and diagnosing chromosomal abnormalities without needing actively dividing cells. It can identify numerical and structural chromosome changes.

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344 views8 pages

Fish Technique: Kirti Shikha

FISH is a cytogenetic technique that uses fluorescent probes that bind to chromosomes to locate specific genes or portions of DNA. It allows researchers to visualize genetic material in individual cells. A probe is made complementary to the target sequence and labeled with a fluorescent marker. The chromosomes are treated so the probe can hybridize to its matching sequence. Under a fluorescent microscope, the probe appears as fluorescent signals, showing where the target sequence is located on the chromosomes. FISH has many applications in research and diagnosing chromosomal abnormalities without needing actively dividing cells. It can identify numerical and structural chromosome changes.

Uploaded by

Saurav Roshan
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPTX, PDF, TXT or read online on Scribd

FISH TECHNIQUE

KIRTI SHIKHA
INTRODUCTION

Fluorescence in situ hybridization (FISH) is a kind of cytogenetic technique which uses fluorescent probes
binding parts of the chromosome to show a high degree of sequence complementarity. Fluorescence
microscopy can be used to find out where the fluorescent probe bound to the chromosome. This technique
provides a novel way for researchers to visualize and map the genetic material in an individual cell,
including specific genes or portions of genes. It is an important tool for understanding a variety of
chromosomal abnormalities and other genetic mutations. Different from most other techniques used for
chromosomes study, FISH has no need to be performed on cells that are actively dividing, which makes it a
very versatile procedure.
Scheme of the principle of the FISH experiment to localize a gene in the nucleus.
TECHNIQUES

FISH is useful, for example, to help a researcher identify where a particular gene falls within an individual's chromosomes.
Here's how it works:
 Make a probe complementary to the known sequence. When making the probe, label it with a fluorescent marker, e.g.
fluorescein, by incorporating nucleotides that have the marker attached to them.
 Put the chromosomes on a microscope slide and denature them.

 Denature the probe and add it to the microscope slide, allowing the probe hybridize to its complementary site.

 Wash off the excess probe and observe the chromosomes under a fluorescent microscope. The probe will show as one or
more fluorescent signals in the microscope, depending on how many sites it can hybridize to.
The five basic steps of FISH. (Oliveira and
French 2005)
FUNCTION

FISH is widely used for several diagnostic applications: identification of numerical and structural
abnormalities, characterization of marker chromosomes, monitoring the effects of therapy, detection of
minimal residual disease, tracking the origin of cells after bone marrow transplantation, identification of
regions of deletion or amplification, detection of chromosome abnormalities in non-dividing or terminally
differentiated cells, determination of lineage involvement of clonal cells, etc. Moreover it has many
applications in research: identification of non-random chromosome rearrangements, identification of
translocation molecular breakpoint, identification of commonly deleted regions, gene mapping,
characterization of somatic cells hybrids, identification of amplified genes, study the mechanism of
rearrangements. FISH is also used to compare the genomes of two biological species to deduce
evolutionary relationships.
TYPES OF F.I.S.H
 Locus specific probes bind to a particular region of a chromosome. This type of probe is useful when
researchers have isolated a small portion of a gene and want to determine on which chromosome the
gene is located.
 Alphoid or centromeric repeat probes are generated from repetitive sequences found in the middle of each
chromosome. Researchers use these probes to determine whether an individual has the correct
number of chromosomes. These probes can also be used in combination with "locus specific probes" to
determine whether an individual is missing genetic material from a particular chromosome.

 Whole chromosome probes are actually collections of smaller probes, each of which binds to a different
sequence along the length of a given chromosome. Using multiple probes labeled with a mixture of
different fluorescent dyes, scientists are able to label each chromosome in its own unique color. The
resulting full-color map of the chromosome is known as a spectral karyotype. Whole chromosome
probes are particularly useful for examining chromosomal abnormalities, for example, when a piece of
one chromosome is attached to the end of another chromosome.
FISH probes Sample

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