Congo Red Assay

Congo Red birefringence assay was developed for the examination of in vivo amyloid and was later applied in the examination of in vitrosamples. Many things other than amyloid exhibit birefringence; buffer salts (phosphate and urea) are inherently birefringent and appear as intense white birefringence, hair or other fibers exhibit birefringence but are often multicolored. This technique is relatively subjective and a known fibrillar sample should be used as a control.

Congo Red spectrophotometric assay is more objective, less prone to misinterpretation, and can be easily combined with the microscopic analysis.

Procedures for Congo Red birefringence assay:

1. Prepare the staining solution: prepare a solution of 80% EtOH: 20% DDI water and add a saturating amount of NaCl. Stir this solution for a few minutes and filter away the excess NaCl. Add a saturating amount of Congo Red, stir, and filter to obtain thefinal working solution. Congo Red can be used straight from the manufacturer or can be recrystallized from 50% EtOH: 50% DDI water prior to use. The staining solution should be used immediately or stored in 0.05% sodiumazide to inhibit bacterial growth.

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