Ex situ and in situ demonstration of amyloid fibrils for confirmation of amyloidosis using transmission electron microscopy.

IF 1.2 3区 农林科学 Q3 VETERINARY SCIENCES
Robert Polon, Christina R Heard, Omar Gonzales-Viera, Melissa Macías-Rioseco, Aslı Mete, Katherine Watson, Leslie W Woods, Aníbal G Armién
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引用次数: 0

Abstract

Confirmation of extracellular amyloid deposition across various animal species and tissue types has been a long-standing challenge in veterinary diagnostic pathology. Transmission electron microscopy (TEM) has historically been used to advance the understanding of amyloid fibril morphology and confirm amyloid fibril deposition when histologic methods provide unclear results. We assessed the feasibility of utilizing TEM for routine confirmation of amyloidosis as an addition to histology. We analyzed ex situ amyloid fibrils with direct, negative-contrast TEM and in situ amyloid fibrils with aldehyde-fixed, plastic-embedding TEM to confirm amyloidosis in a variety of cases in which amorphous extracellular amyloid deposits had been identified by H&E and Congo red staining. We compared the 2 TEM methods and documented amyloid fibril morphology and morphometry in 7 species (goat, guinea pig, kudu, fox, sheep, flamingo, and duck). Ex situ fibrils had helical morphology and widths of 15-18 nm across all species. Fibril crossover distances had more interspecies variation of 60-130 nm, and species could be grouped based on pitch (twist size). Twisting patterns of in situ fibrils could not be visualized, but in situ widths of 10 nm were measured across all species. In 4 different chicken cases, fibrils differing morphologically from amyloid were consistently detected via both TEM methods, suggesting the possibility of a non-amyloid deposit that is commonly diagnosed as amyloidosis based on its histologic appearance. When available, we recommend routine confirmation of amyloid fibril deposition by TEM.

利用透射电子显微镜原位和非原位显示淀粉样纤维,以确认淀粉样变性。
在各种动物物种和组织类型中确认细胞外淀粉样蛋白沉积一直是兽医诊断病理学的长期挑战。透射电子显微镜(TEM)历来被用于促进对淀粉样纤维形态的理解,并在组织学方法提供不明确结果时确认淀粉样纤维沉积。我们评估了利用透射电镜常规确认淀粉样变性的可行性,作为组织学的补充。我们用直接负对比透射电镜分析了非原位淀粉样蛋白原纤维,用醛固定塑料包埋透射电镜分析了原位淀粉样蛋白原纤维,以确认各种病例的淀粉样变性,其中无定形细胞外淀粉样蛋白沉积已被H&E和刚果红染色鉴定。我们比较了两种透射电镜方法,并记录了7种物种(山羊、豚鼠、kudu、狐狸、绵羊、火烈鸟和鸭子)的淀粉样蛋白纤维形态和形态测定。非原位原纤维呈螺旋状,宽度在15 ~ 18 nm之间。纤维交叉距离在60 ~ 130 nm之间有较大的种间差异,可以根据节距(捻度)对物种进行分组。原位原纤维的扭曲模式无法可视化,但在所有物种中都测量了10 nm的原位宽度。在4例不同的鸡病例中,通过两种透射电镜方法一致检测到与淀粉样蛋白形态不同的原纤维,提示可能存在非淀粉样蛋白沉积,根据其组织学外观通常诊断为淀粉样变性。如有可能,我们建议用透射电镜常规确认淀粉样纤维沉积。
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来源期刊
CiteScore
3.00
自引率
6.70%
发文量
127
审稿时长
6-16 weeks
期刊介绍: The Journal of Veterinary Diagnostic Investigation (J Vet Diagn Invest) is an international peer-reviewed journal published bimonthly in English by the American Association of Veterinary Laboratory Diagnosticians (AAVLD). JVDI is devoted to all aspects of veterinary laboratory diagnostic science including the major disciplines of anatomic pathology, bacteriology/mycology, clinical pathology, epidemiology, immunology, laboratory information management, molecular biology, parasitology, public health, toxicology, and virology.
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