Molecular diagnostics is the fastest growing segment of the diagnostic market. Fluorescence in situ hybridization and FISH analysis included a $500 million market for 7 million people. . The FISH market is expected to be 11%, growing to the next five years compared to the same period last year. A market of 4 to 5 million people is produced in the United States. The FISH test is used to identify the form of biomarker DNA/RNA. It is a well-known method for genetic mapping and gene expression analysis. This technique uses a fluorescent dye-labeled probe and maps to a chromosome or frame of core DNA to produce a nucleotide sequence that observes the fluorescent signal in situ by microscopy. It is, for example, a cytospin smear (cell suspension) and then touched with a fresh preparation fixed with a formulation or formalin, or in a paraffin-embedded (FFPE) tissue. It is mainly used for research and clinical diagnosis in the following fields:
• Prenatal diagnosis • Cancer diagnosis • Molecular cytogenetics and mental retardation of birth defects • Chromosomal abnormalities • Clinical genetics • Neuroscience • Reproductive medicine • Toxicology • Microbial ecology • Evolutionary biology • Comparative genomics • Cell genomes Learning • distinguish between benign and malignant tumors
Most of the market's reagents include FDA-approved tests, accompanying diagnostics, research using only detectors, custom reagent probes, and support and supplies. It is necessary to perform hybridization to perform these labor intensive test equipment. Understanding the requirements for hybridization process equipment must be clearly defined.
The necessary equipment is as follows:
- Acid Meter - Balance - Mixing Plate - Micro Centrifuge - Refrigerator - Freezing
Organizational preparation requirements:
- Slicer - Water Bath - Oven
Performing the FISH process will require the following equipment and materials in addition to the reagents:
- Slide Dryer - Water Bath - Hybrid Stove - Coplin Jar - Cover Glass - Dyeing Dish - Rubber Glue (optional: depending on equipment)
- Fluorescence Microscope DAPI Filter (Blue), FITC Filter (Green), Texas Red Filter Photography Capability
FISH process - simplified <br>sample preparation - depariffinization, dehydration, air drying, pretreatment (95 C @ 30 min, Coplin Jar), ​​protease treatment (37 ° C 10 - 20 min, Coplin Jar), ​​dehydration, Air drying, mark, for FISH detectors,
Densification with cement-covered glass - 5 min @ 75 ° C (45 - 50 ° C, medium expansion + additional steps to denature probe) hybridization - 37 ° C, 16 h - 72 h
Labeling process <br> Direct labeling - label binding to the probe and it is less sensitive to indirect labeling - additional steps before detection, with antibodies, results amplified by single detection probe storage - 4 - 8 ° C for up to two months Time, in the dark area - sensitization
Key Process Steps <br> The process of FISH is divided into two steps required for the device to heat the slide; the denaturation step and hybridization. These steps are similar to the most FFPE FISH programs.
Denaturation: denatured probes and double-stranded DNA
Average time: 5 minutes Standard temperature: 75 ° C
Hybridization: Hybridization of probe to target nucleic acid Time range: 16 - 72 hours Standard temperature: 37 ° C
Slides that have been incubated must be stored at 4 - 8 ° C until they are taken under the microscope. They can be stored at these temperatures for several months.
Shanghai Shujun Instrument Equipment Co., Ltd. is the agent of Boekel in situ hybridization products in the United States. For more information, please contact 021-64825207/010-83659476/020-82528560 for consultation or transfer.
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