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.
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that uses fluorescent probes to investigate the presence of small, submicroscopic chromosomal changes that are beyond the resolution of karyotype analysis.
This PowerPoint presentation explain the concept,process and application of Fluorescence insitu hybridization.
Fundamentals of Fluorescence in situ Hybridization Amartya Pradhan
This presentation provides an insight into the fundamentals of in situ hybridization (ISH), especially fluorescence in situ hybridization. It is ideal for classroom lecture.
In situ Hybridization (ISH) and Fluorescence in Situ Hybridization (FISH) Creative-Diagnostics
In situ Hybridization (ISH) and Fluorescence in Situ Hybridization (FISH) by Creative Diagnostics, learn more http://www.creative-diagnostics.com/in-situ-hybridization-and-fluorescence-in-situ-hybridization.htm
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes.
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that uses fluorescent probes to investigate the presence of small, submicroscopic chromosomal changes that are beyond the resolution of karyotype analysis.
This PowerPoint presentation explain the concept,process and application of Fluorescence insitu hybridization.
Fundamentals of Fluorescence in situ Hybridization Amartya Pradhan
This presentation provides an insight into the fundamentals of in situ hybridization (ISH), especially fluorescence in situ hybridization. It is ideal for classroom lecture.
In situ Hybridization (ISH) and Fluorescence in Situ Hybridization (FISH) Creative-Diagnostics
In situ Hybridization (ISH) and Fluorescence in Situ Hybridization (FISH) by Creative Diagnostics, learn more http://www.creative-diagnostics.com/in-situ-hybridization-and-fluorescence-in-situ-hybridization.htm
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes.
This presentation is about DNA fingerprinting, a brief description is given about its principle, working, technique and its application with a example.
What is in situ hybridization
Radioactive ISH
Fluorescent ISH
Colorimetric ISH
ISH: three variables
The sample
The probe
Optimizing ISH Detection
ISH controls
Data Analysis
A cytological technique to detect the nature of adjacent chromosomal regions by using different staining technique assisted with some pre treatment of metaphase chromosomes prepared on the slides
This presentation is about DNA fingerprinting, a brief description is given about its principle, working, technique and its application with a example.
What is in situ hybridization
Radioactive ISH
Fluorescent ISH
Colorimetric ISH
ISH: three variables
The sample
The probe
Optimizing ISH Detection
ISH controls
Data Analysis
A cytological technique to detect the nature of adjacent chromosomal regions by using different staining technique assisted with some pre treatment of metaphase chromosomes prepared on the slides
This powerpoint explains about the nucleic acid hybridization, its principle, application and the assay methods. Also it gives clear picture about DNA probes, its sysnthesis, mechanism of probes and the detector system in DNA hybridization.
To describe DNA extraction
To explain and demonstrate DNA cloning
To explain the process of PCR and its uses.
To explain DNA fingerprinting and its uses
DNA Fingerprinting for Taxonomy and Phylogeny.pptxsharanabasapppa
Deoxyribonucleic acid, a self-replicating material which is present in all living organisms as the main constituent of chromosomes.
DNA is made up of molecules called nucleotides. Each nucleotide contains a phosphate group, a sugar group and a nitrogen base.
The four types of nitrogen bases are adenine (A), thymine (T), guanine (G) and cytosine (C). The order of these bases is what determinesDNA's instructions, or genetic code.
Creative Bioarray offers high quality, time efficient and cost effective kinase solution for assay development, high throughput screening (HTS) and selectivity profiling.
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Creative Bioarray provides an extensive range of high quality RNA samples which are ideal for Northern blotting, ribonuclease protection assay, SI nuclease assay, RT-PCR/Q-PCR analysis, rapid amplification of cDNA ends (RACE) and purification of mRNA for library construction.
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Immunohistochemistry (IHC) is the process of detecting antigens (e.g. proteins) in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues.
https://www.creative-bioarray.com/protocol/immunohistochemistry-protocol.htm
A fluorescent compound has the property of absorbing light energy at a range of specific wavelengths. This absorption of light causes electron to rise from the ground state to a higher energy level (excited state). The excited electron quickly decays to its ground state while releasing the excess energy in the form of photon of light. This transition of energy is called fluorescence.
https://www.creative-bioarray.com/support/fluorochromes-in-flow-cytometry.htm
Creative Bioarray is offering Caco-2 permeability assay to help determine the absorption and the bioavailability of drug candidates, facilitating the lead optimization process in drug discovery.
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Comparison of caco 2 with other cell-based models for intestinal permeability...Creative-Bioarray
The use of cell cultures provides a method to predict drug permeability by utilizing cell monolayers in a two-chamber diffusion system to simulate the passage of drugs from the intestinal lumen into the blood.
Cell cycle refers to the set of events through which a cell grows, replicates its genome, and ultimately divides into two daughter cells through the process of mitosis.
https://www.creative-bioarray.com/cell-cycle-assays.htm
Creative Bioarray is offering Caco-2 permeability assay to help determine the absorption and the bioavailability of drug candidates, facilitating the lead optimization process in drug discovery.
https://www.creative-bioarray.com/Services/caco-2-permeability-assay.htm
A broad range of reprogramming systems is available including mRNA reprogramming, dox-inducible human 4F2A reprogramming and lentivirus reprogramming. These technologies have been adopted and developed in close collaboration with leading iPS cell pioneers. Our extensive range of products allows for customization of individual transcription factors, cell culture conditions, and delivery systems for the optimization of reprogramming parameters.
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Cell proliferation assays are used to monitor the dynamic growth of a cell population or to detect daughter cells in a growing population.
On the other hand, cell viability assays assess how healthy the cells are by measuring markers of cellular activity.
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By using flow cytometry, staining dyes are needed. Creative Bioarray can choose different dyes to perform the assays, including propidium iodide (PI), BrdU, 7-amino actinomycin-D (7-AAD), Hoechst 33342 and 33258, and 4’6’-diamidino-2-phenylindole (DAPI), based on the customer’s applications or requirements.
https://www.creative-bioarray.com/cell-cycle-assays.htm
By using flow cytometry, staining dyes are needed. Creative Bioarray can choose different dyes to perform the assays, including propidium iodide (PI), BrdU, 7-amino actinomycin-D (7-AAD), Hoechst 33342 and 33258, and 4’6’-diamidino-2-phenylindole (DAPI), based on the customer’s applications or requirements.
https://www.creative-bioarray.com/cell-cycle-assays.htm
Induced Pluripotent Stem Cells (iPSCs) are a type pf pluripotent stem cell artificially derived, and often referred to as programmed, from adult somatic cells using the expression of certain genes in culture.
https://www.creative-bioarray.com/products/ipsc-reprogramming-kit-list-239.htm
A fluorescent compound has the property of absorbing light energy at a range of specific wavelengths. This absorption of light causes electron to rise from the ground state to a higher energy level (excited state). The excited electron quickly decays to its ground state while releasing the excess energy in the form of photon of light. This transition of energy is called fluorescence.
https://www.creative-bioarray.com/support/fluorochromes-in-flow-cytometry.htm
Creative Bioarray provides our global clients the most comprehensive histology services. Drawing on many years of experience and in-depth knowledge, Creative Bioarray offers tissue processing, embedding, sectioning, and staining. Besides a histological examination of all major organs/tissues is provided, including immunohistochemistry (IHC), immunofluorescence (IF), in situ hybridization (ISH), fluorescent in situ hybridization (FISH), and transmission electron mircoscopy.
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Creative Bioarray offers 35 human cell systems with over 160 different cell types. Moreover, we also provide our customers with primary cells from over 13 types of other animals.
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Drug transporters mediate the uptake and efflux of a broad variety of drugs and drug metabolites. Most uptake transporters are members of the solute carrier (SLC) family, while most efflux transporters are members of the ATP-binding cassette (ABC) transporter family.
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Acroscell provides ready-to-use beating human induced pluripotent stem cells (iPSC)-derived cardiomyocytes. Generated from mature cells that have been genetically reprogramed to a pluripotent stem cell state, induced pluripotent stem cells (iPSCs) can be readily expanded and induced to specialize or differentiate into cardiomyocytes in vitro.
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PAMPA provides a simplified approach to permeability by addressing just a single transport mechanism. This avoids the complexities of active transport and metabolism, enabling ranking of the compounds on a single permeability factor.
https://dda.creative-bioarray.com/parallel-artificial-membrane-permeability-assay.html
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
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. 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. 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.
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. 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
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. 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
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. 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. For more info please contact us:
E-mail: info@creative-bioarray.com
Go to our website：
www. creative-bioarray.com
Editor's Notes
Here are four parts. What is FISH? The probes ,the procedure of FISH and the applications.
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.
The DNA contains two strand-like molecules coiled together into a structure known as a double helix. The bases in each strand are able to bind to each other and hold the DNA together. When two complementary sequences find each other they will bind together, or hybridise. FISH works by exploiting the ability of one DNA strand to hybridise specifically to another DNA strand.
FISH works by exploiting the ability of one DNA strand to hybridise specifically to another DNA strand.FISH uses small DNA strands called probes that have a fluorescent label attached to them. The probes are complementary to specific parts of a chromosome. When DNA is heated, the patient’s two DNA strands break apart, or denature, and the probes are able to hybridise to their complementary sequence in the patient’s DNA. If a small deletion is present in the region complementary to the probe, the probe will not hybridise. If a duplication is present, more of the probe is able to hybridise.
Probe is critical to in situ hybridization, and a right probe can help you achieve your goals. Not only the probe types but also the label of probe should you take into account when you choose a probe for in situ hybridization.
Different probes have different characteristics. dsDNA probes are stable and easier to obtain. ssDNA probes are more specific and better in resistanting to Rnases. For RNA probes, they have higher thermal stability and better tissue penetration.
There are four steps of FISH:Preparation of the fluorescent probes; Denaturation of the probe and the target; Hybridization of the probe and the target; Detection.
The samples of FISH are two categories: fixed cell suspension; Formalin fixed paraffin embedded tissues. Sample predation includes sample fixation and slides.
Probe is critical to in situ hybridization, and a right probe can help you achieve your goals. Not only the probe types but also the label of probe should you take into account when you choose a probe for in situ hybridization.
When you get the slides，the next steps are dehydration and hybridization.
And the last step is detection。Fluorescence in situ hybridization, as an important biological experimental technique, it features in situ and without PCR, and can be used for quantitative analysis of specific microorganisms in environmental samples.
FISH can be used for different areas，such as morphology，pathology，developmental biology，and karyotyping and phylogenetic analysis.
Creative Bioarray provides comprehensive commercial probes and FISH Kits for easy to use. In addition, high-quality services are also available for our clients.
