Diagnosis of renal amyloidosis using Congo red fluorescence

Şen, Sait; Başdemir, Gülçin

doi:10.1046/j.1440-1827.2003.01513.x

Cited by 32 publications

(34 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Upon binding to amyloid fibrils, CR gives a characteristic applegreen birefringence (or perhaps more correctly, shows anomalous colors) when examined between crossed polarizer and analyzer, and a characteristic shift in absorbance maximum from about 490 nm to about 540 nm is seen [32][33][34][35][36]. CR is most often applied in diagnosis of amyloid in ex vivo tissue sections using polarization microscopy [2], but light microscopy [37] and fluorescence microscopy [25,37] have also been used. In vitro quantification of insulin or amyloid β (Aβ) amyloid upon CR binding can be performed using absorption spectroscopy [38,39], although CR lacks sensitivity at low amyloid fibril concentration compared to ThT [40].…”

Section: Overview Of Molecular Probes For Amyloid Fibril Detectionmentioning

confidence: 99%

Binding mode of Thioflavin T and other molecular probes in the context of amyloid fibrils—current status

2009

View full text Add to dashboard Cite

Because understanding amyloid fibrillation in molecular detail is essential for development of strategies to control amyloid formation and overcome neurodegenerative disorders, increased understanding of present molecular probes as well as development of new probes are of utmost importance. To date, the binding modes of these molecular probes to amyloid fibrils are by no means adequately described or understood, and the large number of studies on Thioflavin T (ThT) and Congo Red (CR) binding have resulted in models that are incomplete and conflicting. Different types of binding sites are likely to be present in amyloid fibrils with differences in binding modes. ThT may bind in channels running parallel to the long axis of the fibril. In the channels, ThT may bind in either a monomeric or dimeric form of which the molecular conformation is likely to be planar. CR may bind in grooves formed along the β-sheets as a planar molecule in either a monomeric or supramolecular form.

show abstract

Section: Overview Of Molecular Probes For Amyloid Fibril Detectionmentioning

confidence: 99%

Binding mode of Thioflavin T and other molecular probes in the context of amyloid fibrils—current status

2009

View full text Add to dashboard Cite

show abstract

“…In addition, compared to section thicknesses of 4-6 μm used in most studies so far [7,15,16], we could demonstrate that using thinner sections of 2 μm, which are advantageous for immunohistochemistry (better signal-to-noise ratio, less detachment artifacts), can also lead to satisfactory results regarding Congo red staining, if read out by a fluorescent microscope.…”

Section: Discussionmentioning

confidence: 74%

“…Several earlier studies have focused on the combination of fluorescence and immunohistochemistry [7,15,16]. Here, we present additional evidence for the practicability of such an approach and report a protocol easily translatable into routine, which enabled us to accurately address amyloid subtypization on formalin-fixed and paraffin-embedded tissues.…”

Section: Introductionmentioning

confidence: 80%

A More Accurate Approach to Amyloid Detection and Subtyping: Combining in situ Congo Red Staining and Immunohistochemistry

Menter

Bachmann²,

Grieshaber³

et al. 2016

Pathobiology

View full text Add to dashboard Cite

Amyloidosis is the result of various, differently approachable diseases. It is vital to subtype the amyloid deposits in order to establish and finally treat the underlying disease properly. Besides the classical staining with Congo red, further procedures like immunohistochemical staining are needed for classification. Here, we present a more accurate approach using Congo red/immunohistochemical double staining easily applicable in routine diagnostics. Modifications of the Congo red staining technique and the immunohistochemical procedures were needed in order to combine both staining procedures on one slide. The evaluation was done using conventional light and fluorescence microscopy. By shortening the staining time for Congo red to 10 s and by modification regarding endogenous peroxidase blockage, accurate results could be obtained for evaluating the Congo red/immunohistochemistry double staining using a fluorescence microscope. Sections of 2 μm instead of 4 μm thickness were superior for evaluation, since they increased staining specificity. The combination of Congo red and immunohistochemistry as in situ double staining on one slide is a feasible approach in the diagnosis of amyloidosis. It allows focusing on the fluorescent Congo red-positive areas when evaluating immunohistochemistry, thus avoiding signing out false-positive results. Additionally, it increases the signal-to-noise ratio of the immunohistochemically stained sections on conventional microscopy.

show abstract

“…However, some difficulties of the CR staining method such as requiring a thick section reaching 12 micron thickness and optimization of the staining method are known (1,10,11). Thioflavin S and T, the metachromatic stains of toluidine, methyl or crystal violet as well as CR fluorescence are ancillary methods in amyloidosis diagnosis (10,11,(13)(14)(15). Amyloid stains are generally used in renal biopsies in our country while they are only used if there is a clinical and pathological suspicion of amyloid deposits in other organ biopsies.…”

Section: Discussionmentioning

confidence: 99%

Digitally Reinforced Polarization of Hematoxylin-Eosin in the Diagnosis of Renal Amyloidosis

Şen

Kumbaraci²

2012

TJPATH

Self Cite

View full text Add to dashboard Cite

Objective: Systemic amyloidosis is a rare disorder, characterized by extracellular accumulation of Congo red positive fibrillar amyloid protein deposits that have an amorphous, eosinophilic appearance on hematoxylin-eosin stained preparations. The kidney is the most commonly affected organ by systemic amyloidosis. Congo red staining increases the positive birefringence of the weakly birefringent unstained amyloid. In this study, we investigated the potential diagnostic power of digitally reinforced birefringence of routine hematoxylin-eosin stained slides from renal biopsies. Material and Method:We reviewed 130 hematoxylin-eosin stained slides for polarization. Sixty-five new amyloidosis cases were diagnosed by renal biopsy. All renal biopsies were evaluated by light microscopy and immunofluorescence. Slides were reevaluated blindly using a microscope (Olympus BX51) that was attached polarization filters and connected to a digital camera (Olympus DP21, SAL). Deposits that showed green birefringence on hematoxylin-eosin preparations with digitalized microscopy were considered positive and the results were confirmed using Congo red.Results: Of the 65 Congo red confirmed amyloid positive biopsies, 61 showed green birefringence with hematoxylin-eosin. Of the 65 Congo-red confirmed amyloid negative biopsies, two were considered to be false positive. The sensitivity, specificity, and positive and negative predictive values were estimated as 94%, 97%, 97% and 94% respectively. Conclusion:We concluded that polarized hematoxylin-eosin sections can be used digitally as a fast and first step diagnostic method for renal amyloidosis Key Words: Amyloid, Amyloidosis, Congo Red, Hematoxylin ÖZ Amaç: Sistemik amiloidozlar, hematoksilen-eozin boyamada amorf eozinofilik görülen, Kongo kırmızısı ile boyanan fibriller amiloid proteinlerin ekstrasellüler birikimiyle karakterize nadir hastalıklardır. Böbrekler sistemik amiloidozlardan en sık etkilenen organdır. Kongo kırmızısı, zayıf birefrenjant boyanmamış amiloidin birefranjansını artırır. Bu çalışmada, böbrek biopsilerinin rutin hematoksilen eozin kesitlerinde dijital güçlendirilmiş birefrenjansın potansiyel tanısal gücünü araştırdık. Gereç ve Yöntem:Hematoksilen-eozin boyalı 130 preparat polarizasyon için değerlendirildi. Altmış beş yeni amiloidoz olgusuna böbrek biyopsisi ile tanı konuldu. Tüm böbrek biopsileri ışık ve immünflöresan mikroskop ile değerlendirildi. Preparatlar kör olarak, polarizasyon filtreleri eklenmiş ve digital kamera (Olympus DP21, SAL) bağlanmış mikroskopla (Olympus BX51) değerlendirildi. Dijitalize mikroskopi ile yeşil birefrenjans gösteren birikimler pozitif olarak değerlendirildi ve Kongo kırmızısı ile birikimler doğrulandı. Bulgular: Kongo kırmızısı ile doğrulanmış 65 amiloid pozitif biopsinin 61'i hematoksilen-eozin ile yeşil birefrenjans gösterdi. Kongo kırmızısı ile doğrulanmış, 65 amiloid birikimi olmayan olgunun 2'si yalancı pozitif olarak değerlendirildi. Duyarlılık, özgüllük, pozitif ve negatif prediktif değerler sırasıyla %94, %97, %97 ve ...

show abstract

Diagnosis of renal amyloidosis using Congo red fluorescence

Cited by 32 publications

References 24 publications

Binding mode of Thioflavin T and other molecular probes in the context of amyloid fibrils—current status

Binding mode of Thioflavin T and other molecular probes in the context of amyloid fibrils—current status

A More Accurate Approach to Amyloid Detection and Subtyping: Combining in situ Congo Red Staining and Immunohistochemistry

Digitally Reinforced Polarization of Hematoxylin-Eosin in the Diagnosis of Renal Amyloidosis

Contact Info

Product

Resources

About