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Fluorescent In Situ Hybridization (FISH) Assay
Content
Definition, Principle and Sample Types
What is FISH?
The core of FISH technology
Probes
A quick and simple FISH pr...
输入标题文本
What is Fluorescent in situ hybridization (FISH) ?
• Fluorescent in situ hybridization (FISH) is a molecular
cytoge...
输入标题文本
FISH works by exploiting the ability of one DNA strand to
hybridise specifically to another DNA strand.
How does FI...
输入标题文本
heating
probe with
fluorescent label
hybridisation
denaturation
cooling
FISH Technology
输入标题文本
Centromere probes
• Alpha and Satellite III probes
• Generated from repetitive sequences
found in centromeres
• Cen...
输入标题文本
dsDNA probes
Stable, available, easier to obtain Stable, easier to work with, more
specific, resistant to RNases,
b...
输入标题文本
• 32P
• 35S
• 3H
• Biotin
• Digoxigenin
• Fluorescent dy
Probes Labeling
输入标题文本
Denaturation of the
probe and the target
Hybridization of the
probe and the target
Preparation of the
fluorescent p...
输入标题文本
Sample Types
• Fixed cell suspension
• Formalin fixed paraffin embedded tissues
The Sample Preparation and Sample T...
输入标题文本
Preparation of the fluorescent probes
• Commercial Probes can be provided by many
biotech companies
• Based on your...
输入标题文本
Denaturation of the probe and the target
Dehydration
(1) Place the suitable amount of fixed sample on the slide,
(2...
输入标题文本
Detection by Microscope
输入标题文本
1Morphology Morphology and population structure of microorganisms
2Patholog
y
Pathogen profiling, abnormal gene exp...
输入标题文本
Products
FISH Services
In addition to products, high-quality services are also
available for our clients.
We provid...
For more info please contact us:
E-mail: info@creative-bioarray.com
Go to our website:
www. creative-bioarray.com
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Fluorescent in situ hybridization (fish) assay

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Fluorescent in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome with a high degree of sequence complementarity. Creative Bioarray provides comprehensive FISH services and products to our clients.

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Fluorescent in situ hybridization (fish) assay

  1. 1. Fluorescent In Situ Hybridization (FISH) Assay
  2. 2. Content Definition, Principle and Sample Types What is FISH? The core of FISH technology Probes A quick and simple FISH protocol FISH Procedure Wide range of uses and bright prospects Application 1 2 3 4
  3. 3. 输入标题文本 What is Fluorescent in situ hybridization (FISH) ? • Fluorescent in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome with a high degree of sequence complementarity. • It is used to detect and localize the presence or absence of specific DNA sequences on chromosomes. Definition
  4. 4. 输入标题文本 FISH works by exploiting the ability of one DNA strand to hybridise specifically to another DNA strand. How does FISH work ? The Structure of DNA The DNA contains two strand-like molecules coiled together into a structure known as a double helix. The principle of complementary base pairing When two complementary sequences find each other they will bind together, or hybridise.
  5. 5. 输入标题文本 heating probe with fluorescent label hybridisation denaturation cooling FISH Technology
  6. 6. 输入标题文本 Centromere probes • Alpha and Satellite III probes • Generated from repetitive sequences found in centromeres • Centromere regions are stained brighter • Collection of probes that bind to the whole length of chromosome • Multiple probe labels are used • Hybridize along the length of the chromosome • Specific for telomeres • Specific to the 300 kb locus at the end of specific chromosome • Deletion • Translocation probes • Gene detection & localization probes • Gene amplification probes Telomere Whole chromosome Locus Probes What Kind of Probes Can Be Used?
  7. 7. 输入标题文本 dsDNA probes Stable, available, easier to obtain Stable, easier to work with, more specific, resistant to RNases, better tissue penetration, without self- hybridize • Higher thermal stability, • Better tissue penetration, • More specific, • Low background noise by RNase •Economical, stable, available, easier to work with, •more specific, resistant to RNases, •better tissue penetration, better reproducibility. RNA probes ssDNA probes Synthetic oligonucleotides probes Probes Characteristics of Different Probes Types
  8. 8. 输入标题文本 • 32P • 35S • 3H • Biotin • Digoxigenin • Fluorescent dy Probes Labeling
  9. 9. 输入标题文本 Denaturation of the probe and the target Hybridization of the probe and the target Preparation of the fluorescent probes Protocol Outline
  10. 10. 输入标题文本 Sample Types • Fixed cell suspension • Formalin fixed paraffin embedded tissues The Sample Preparation and Sample Types Preparation • Sample fixation • Slide tissue
  11. 11. 输入标题文本 Preparation of the fluorescent probes • Commercial Probes can be provided by many biotech companies • Based on your special needs, custom probes are also synthesized • e.g. 𝒂-satellite DNA is often chosen as the source of centromeric probes.
  12. 12. 输入标题文本 Denaturation of the probe and the target Dehydration (1) Place the suitable amount of fixed sample on the slide, (2) put into 46°C oven drying for 10 minutes (3) Followed by immersion 50%, 80%, 96% ethanol solution, each for 3 minutes (4) dry in the air Hybridization (1) Add 10 μl of Hybridization buffer to the sample on the glass plate and try to cover the entire sample (2) Add 1 μl probe to Hybridization buffer (3) Put a wet absorbent paper in a 50ml centrifuge tube, put the glass pieces in, then place them in a 46°C incubator for 1.5 hours (4) Preheat the Washing buffer to 48°C for the next step
  13. 13. 输入标题文本 Detection by Microscope
  14. 14. 输入标题文本 1Morphology Morphology and population structure of microorganisms 2Patholog y Pathogen profiling, abnormal gene expression 3 Developmental biology Gene expression profiling in embryonic tissues 4 Karyotyping and phylogenetic analysis Unique FISH patterns on individual chromosomes, chromosomal aberrations Applications
  15. 15. 输入标题文本 Products FISH Services In addition to products, high-quality services are also available for our clients. We provide comprehensive commercial probes and FISH Kits for easy to use. Creative Bioarray
  16. 16. For more info please contact us: E-mail: info@creative-bioarray.com Go to our website: www. creative-bioarray.com
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Fluorescent in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome with a high degree of sequence complementarity. Creative Bioarray provides comprehensive FISH services and products to our clients.

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