Abstract
Recently, fluorescence microscopy becomes more available, presenting new opportunities to face several challenges of experimental biology and medicine. The study was aimed to assess the effectiveness of fluorescence microscopy for the identification of amyloid deposits in human tissues. Post-mortem samples of the myocardium (n = 12) and cerebral cortex (n = 8) obtained from subjects of both sexes aged 60–98 with verified amyloidosis were used as a material for the study. The specimens were stained using 11 different histochemical dyes and subsequently analyzed by light and fluorescence microscopy. Qualitative and quantitative analysis has shown that Thioflavin T is the most effective stain for fluorescence detection of β- and transthyretin amyloid in human tissues. Congo red staining is highly effective for the detection of transthyretin amyloidosis, however, it is ill-suited for the identification of β-amyloid plaques. It has been found that the ability of Congo red to exhibit fluorescence when binding to amyloid fibrils can be used for verification of amyloid deposits instead of the traditional polarized light microscopy. As has been first noted, methyl violet can selectively bind to β-amyloid with fluorescent complex formation. In addition, methyl violet treatment effectively reduces the autofluorescent background in the nervous tissue. This makes methyl violet staining a promising diagnostic tool for Alzheimer's-type pathology.
Highlights
Fluorescence microscopy becomes more available, presenting new opportunities to face several challenges of experimental biology and medicine
Three dyes are capable of binding to amyloid fibrils to form fluorescent complexes: Thioflavin T (ThT), Congo red and methyl violet
Amyloid plaques are characterized by morphological heterogeneity: some plaques have a roundish intensely fluorescent dense central core surrounded by a fibrillar peripheral halo (Fig. 1А; arrow 1), while
Summary
Fluorescence microscopy becomes more available, presenting new opportunities to face several challenges of experimental biology and medicine. The study was aimed to assess the effectiveness of fluorescence microscopy for the identification of amyloid deposits in human tissues. Qualitative and quantitative analysis has shown that Thioflavin T is the most effective stain for fluorescence detection of β- and transthyretin amyloid in human tissues. Целью настоящей работы было оценить эффективность применения метода флуоресцентной микроскопии для идентификации амилоидных скоплений в тканях человека. Проведенные качественный и количественный анализ показали, что тиофлавин Т является наиболее эффективным красителем для флуоресцентного выявления β- и транстиретинового амилоида в тканях человека. Методика окраски конго красным обладает высокой эффективностью в отношении транстиретинового амилоидоза, но плохо подходит для идентификации β-амилоидных бляшек. Установлено, что способность конго красного флуоресцировать при связывании с амилоидными фибриллами может быть использована для верификации амилоидных скоплений вместо традиционной поляризационной микроскопии. А. Суфиева — количественный анализ данных; Д. Л. Цыба — количественный анализ данных; Д. Соблюдение этических стандартов: исследование проведено в соответствии с требованиями Хельсинской декларации Всемирной медицинской ассоциации (2013) и одобрено этическим комитетом ФГБНУ «ИЭМ» (протокол No 3/18 от 22 ноября 2018 г.)
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