Fluorescent staining of frozen sections for amyloid (AMYLOID) thioflavine T

Summary

Fluorescent staining of protein fibers

Principle

Thioflavine T, a cationic benzothiazole derivative, is the most commonly used amyloid fluorescent probe for in vivo or in vitro recognition of fibrosis kinetics and quantitative analysis by methylation of dehydrothiotoluidine in the presence of hydrochloric acid. Compared to thioflavine S, thioflavine T is more stable and broad-spectrum, which makes it better for in vivo detection, especially in the liquid state. Once bound to amyloid fibrils (oligomers), its fluorescence intensity (excitation wavelength 440nm, emission wavelength 490nm), is significantly enhanced, showing apple green fluorescence.

Appliance

Starch. Peptide research

Operation method

Fluorescent staining of frozen sections for amyloid (AMYLOID) thioflavine T

Materials and Instruments

【Materials】Frozen tissue section; frozen section amyloid (AMYLOID)
Reagents and kits]: Thioflavine T (thioflavine T), fluorescence staining kit

Move

Remove 10 frozen tissue sections to be tested.Carefully add xx microliters of YIJI Cleaning Solution (Reagent A) to the slides, spreading it over the entire surface of the slides.Carefully remove the YIJI Cleaning Solution (Reagent A) from the sections.Carefully add xx microliters of YIJI Fixative (Reagent B) to the slides, spreading it over the entire surface of the slides.Incubate in refrigerator at 4 for 10 minutesCarefully remove the YIJI Fixative (Reagent B) from the sections.Carefully add xx microliters of YIJI Cleaning Solution (Reagent A) to the slice, spreading it over the entire surface of the slice sample.Carefully remove the YIJI Cleanup Solution (Reagent A) from the section.Carefully add xx microliters of YIJI Staining Solution (Reagent C) and spread over the entire surface of the section.Incubate at room temperature for 10 minutes, avoiding light.Carefully remove the YIJI Stain (Reagent C) from the sections.Carefully add xx microliters of YIJI Cleaning Solution (Reagent A) to the section and spread over the entire surface of the section.Incubate for 1 minute at room temperatureCarefully remove the YIJI Cleaning Solution (Reagent A) from the sections.Carefully add xx microliters of YIJI Liquid Strengthening Solution (Reagent D) and spread over the entire surface of the slice sample.Incubate for 10 minutes at room temperature Carefully remove the YIJI Reagent (Reagent D) from the sections.Carefully add xx microliters of YIJI Cleaning Solution (Reagent A) to the slice and spread over the entire surface of the slice sample.Incubate for 3 minutes at room temperatureCarefully remove the YIJI Cleaning Solution (Reagent A) from the sections.Seal the sliceImmediately observe under a (confocal) fluorescence microscope (excitation wavelength 440nm, emission wavelength 490nm):Amyloid: apple green fluorescence

Caveat

This product is for 50 operationsWear gloves when handlingThe reagent is corrosive, pay attention to the safety of operationAvoid air bubbles when the reagent solution is on the surface of the sample, and make sure it is spread over the surface of the sample.Fluorescence microscope observation immediately after staining of tissue cells Quercetin, curcumin, resveratrol, etc. may interfere with the assay.

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