Congo Red Method.
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Congo Red Stain * Component test codes cannot be used to order tests. Add 5ÂµL of the Congo Red solution to the phosphatebuffer, scan between 400 and 700 nm and recordthe spectrum. nCounter PanCancer Immune Profiling Panel, DNA Hybridization Capture Target Enrichment, BCR-ABL Negative Myeloproliferative Disease, Next-Generation Sequencing (NGS) for Myeloid and Suspected Myeloid Malignancies, Next-Generation Sequencing (NGS) for Solid Tumors, Precision Medicine in the COVID-19 Crisis, A formalin-fixed, paraffin-embedded (FFPE) tissue block is preferred specimen type, One (1) unbaked, unstained slide cut at 4-5 microns for H&E staining (required) and two to three (2-3) positively charged unstained slides cut at 10 microns.
Examine the stained sample using polarized light microscopy.The microscope should have high quality,strain-free lenses for optimal performance. Clean off excess permount by dipping in xylene solution. Correct CPT coding is the sole responsibility of the billing party. (fat sample never really dries in this time, but adheres better if you do this). Rinse in distilled water. Due to a color change from blue to red at pH 3.0–5.2, Congo red can be used as a pH indicator. Test Description. 3:358–385. Since this color change is an approximate inverse of that of litmus, it can be used with litmus paper in a simple parlor trick: add a drop or two of Congo red to both an acid solution and a base solution. Apple-green birefringence of Congo red stained preparations under polarized light is indicative of the presence of amyloid fibrils. All rights reserved. Dehydrate, clear and mount.
Congo Red birefringence assay was developed for the examination of in vivo amyloid and was later applied in the examination of in vitro samples. Mathematically subtract the Congo Red spectrum from the protein+ Congo Red spectrum. Congo Red birefringence assay was developed for the examination of in vivo amyloid and was later applied in the examination of in vitro samples. The reason for this is that the Congo red dye used in the stain is highly selective for amyloid deposits, which can lead to amyloidosis and also alzheimers. To have a better experience, please upgrade to Microsoft Edge, Google Chrome, or Firefox. 27:643–657. At this stage, a red precipitate may become visible.
Although reportedly first used to stain human tissue in 1886 (see H. Greisbach), it would not be until 1922 that the dye’s specificity for amyloid would be exploited (see H. Bennhold). However, the drawbacks and artefacts obtained when using this dye can be found … More recently, Clement and Truong have shown how fluorescence microscopy using a Texas red filter may enhance the identification of amyloid deposits in Congo red-stained tissue sections, especially when deposits are small. Procedures for Congo Red birefringence assay: 1. An evaluation of Congo red fluorescence for the diagnosis of amyloidosis. Fix slides in formalin (under hood) for at least.
Congo Red (CR) Staining – Fat Pad Biopsies Label slides with patient name. PubMed PMID: 11175644. David P. Steensma, MD, offers readers a fascinating history of the Congo red stain (see references), which still remains the most widely used and cost-effective way to identify amyloid deposits in pathology specimens. The ability to detect amyloid deposits in human tissue is the critical first step in making a diagnosis of amyloidosis. 4. This condition of deposition of amyloid in tissues is known as Amyloidosis. Prepare the staining solution: prepare a solution of 80% EtOH: 20% DDI water and add a saturating amount of NaCl. 2014 Aug;45(8):1766-72. Amyloidosis is a rare disease characterized by the deposition of insoluble misfolded proteins in various tissues and organs. All rights reserved. Additionally, Congo red is used for the diagnostics of the Shigella flexneri serotype 2a, where the dye binds the bacterium's unique lipopolysaccharidestructure. 6. The ability to detect amyloid deposits in human tissue is the critical first step in making a diagnosis of amyloidosis. © 2020 LaboratoryInfo.com. AGFA marketed the dye under the name "Congo red", a catchy name in Germany at the time of the 1884 Berlin West Africa Conference, an important event in the Colonisation of Africa. Place coverslips on slides using permount solution. The information provided here is not sufficient for interface builds; for a complete test mix, … 8. Place in 95%, 95%, 100%, 100% alcohol solutions respectively followed by two separate xylene rinses (dip repeatedly until slides are completely bathed, blot bottom of rack with paper towel to remove excess solution before proceeding to the next rinse). It is still used in histology to stain tissues for microscopic examination, and to serve as an … Add 5â10ÂµL of protein solution to the cuvette and incubate for 30 min at room temperature. If you delete your browser cookies, you will need to rebuild your test list. Fort Myers, FL 33913 He was looking for textile dyes that did not require a mordant step.
Wait for seconds and then blot away the excess solution with a lint-free Kimwipe(or hardened filter paper), being careful not to touch the sample, and allow the stained sample to dry at room temperature. T: 239.768.0600 Griesbach, H. Weitere untersuchungen über azofarbstoffe behufs tinction menschlicher und thierischer gewebe [Further studies of azo dyestuffs as a stain of human and animal tissues]. Congo Red Stain * Component test codes cannot be used to order tests. Air-dry the sample and analyze under polarized light. 3. It is an azo dye.
Deparaffinize and bring sections to distilled water. Stain in congo red working solution for 10 minutes.
Congo Red spectrophotometric assay is more objective, less prone to misinterpretation, and can be easily combined with the microscopic analysis. Add a saturating amount of Congo Red, stir, and filter to obtain thefinal working solution. Air-dry 10ÂµL of the aggregated protein solution onto a glass microscope slide. Congo red staining detects amyloid deposits in brain tissue of amyloid precursor protein transgenic mice and human Alzheimer's tissue. Make sure cold pack is not in direct contact with specimen. Zero a UVâVis spectrophotometer between 400 and700 nm at room temperature with a sample of 1mL phosphate buffer in a disposable cuvette. © Boston University. Procedures for Congo Red birefringence assay: 1. David P. Steensma, MD, offers readers a fascinating history of the Congo red stain (see … Differentiate quickly (9-10 dips) in alkaline alcohol solurion. As you browse our test menu, and you see your preferred test, simply select the Add to Favorites button.
5. Congo Red is the sodium salt of benzidinediazo-bis-1-naphthylamine-4-sulfonic acid; a diazo dye that is red in alkaline solution and blue in acid solution and used especially as an indicator and as a biological stain. Congo red dye forms nonpolar hydrogen bonds with amyloid and red to apple green birefringence occurs when viewed by polarized light due to alignment of dye molecules on the lineraly arranged amyloid fibrils. Congo red has a propensity to aggregate in aqueous and organic solutions. 12701 Commonwealth Dr., Suite 9 In histology and microscopy, Congo red is used for staining in amyloidosis, and for the cell walls of plants and fungi, and for the outer membrane of Gram-negative bacteria.
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