thioflavin s solution  (Millipore)

Journal: Molecular Metabolism

Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

doi: 10.1016/j.molmet.2017.05.016

Figure Lengend Snippet: IL-1Ra immunoreactivity is detectable in human islet α-cells. (A) Paraffin-embedded sections from human islets transduced with Ad-prohIAPP-siRNA (or Ad-control-siRNA) and cultured (11.1 mmol/L glucose) for 7 days were immunolabeled for glucagon (red) and IL-1Ra (green). (B) Triple immunostaining of pre-culture human islets for insulin (blue), glucagon (red) and IL-1Ra (green). The squares (dashed white lines) denote regions enlarged and depicted as inserts at the bottom right of the corresponding images. Amyloid formation during islet culture is shown as an insert in merged micrographs (top right, insulin; red and thioflavin S; blue). Scale bar = 50 μm; inserts: ×3. Micrographs are representative of four independent studies (4 human islet preparations).

Article Snippet: For thioflavin S staining, sections were incubated with 0.5% (wt/vol.) thioflavin S solution (Sigma–Aldrich, Oakville, ON, Canada) for 5 min at room temperature.

Techniques: Transduction, Cell Culture, Immunolabeling, Triple Immunostaining

Journal: Molecular Metabolism

Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

doi: 10.1016/j.molmet.2017.05.016

Figure Lengend Snippet: IL-1Ra levels are increased in oligomer-positive islet areas and reduced in thioflavin S-positive islet areas. (A) Paraffin embedded sections from 7-day cultured (11.1 mmol/L glucose) non-transduced or transduced with Ad-prohIAPP-siRNA (or Ad-control-siRNA) islets were immunolabeled for insulin (red) and IL-1Ra (green). The squares (dashed white lines) denote enlarged regions shown as inserts. (B) Human islets were immunolabeled for IL-1Ra (green) and A11 (red; top panel) or insulin (red), IL-1Ra (green) and thioflavin S (Thio S; blue; bottom panel). (C) Paraffin-embedded sections from (left to right): ob/ob mouse and human adipose tissue immunolabeled for IL-1Ra (green; positive control); human islets incubated with secondary antibody alone (negative control) and anakinra-treated human islets immunolabeled for insulin (red) and IL-1Ra (green). Scale bar = 50 μm; inserts: ×3 (A11: ×4). Micrographs are representative of four independent studies (4 human islet preparations).

Article Snippet: For thioflavin S staining, sections were incubated with 0.5% (wt/vol.) thioflavin S solution (Sigma–Aldrich, Oakville, ON, Canada) for 5 min at room temperature.

Techniques: Cell Culture, Transduction, Immunolabeling, Positive Control, Incubation, Negative Control

Journal: Molecular Metabolism

Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

doi: 10.1016/j.molmet.2017.05.016

Figure Lengend Snippet: Treatment with neutralizing IL-1β antibody markedly reduces islet IL-1β immunoreactivity and prevents β-cell Fas upregulation induced by amyloid formation in cultured human islets. Human islets non-transduced or transduced with Ad-prohIAPP-siRNA or Ad-control-siRNA (MOI: 20) were cultured with (+nIL1β) or without (-nIL1β) a neutralizing monoclonal human IL-1β antibody (1 μg/ml) in CMRL (11.1 mmol/L glucose) for 7 days. (A) Paraffin-embedded islet sections were immunolabeled for insulin (red), IL-1β or Fas (green) and thioflavin S (Thio S; blue) as indicated. The squares (dashed white lines) denote regions enlarged and depicted as inserts at the bottom right of each image. The top right inserts in each micrograph show immunolabeling for insulin (green) and A11 (red). Scale bar = 50 μm; inserts: ×3 (A11: ×4). (B) The proportion of Fas positive islet β-cells (fold over day 0). The percentage of (C) A11 (oligomer)-positive islets, (D) thioflavin S (amyloid) positive islets, and (E) amyloid area to total islet area. Data are presented as mean ± SEM of four independent studies (4 human islet preparations); n = 15–20 islets per condition in each study. *vs day 0; #vs corresponding non-treated group ( p

Article Snippet: For thioflavin S staining, sections were incubated with 0.5% (wt/vol.) thioflavin S solution (Sigma–Aldrich, Oakville, ON, Canada) for 5 min at room temperature.

Techniques: Cell Culture, Transduction, Immunolabeling

Journal: Journal of Cardiovascular Magnetic Resonance

Article Title: Hemorrhage promotes inflammation and myocardial damage following acute myocardial infarction: insights from a novel preclinical model and cardiovascular magnetic resonance

doi: 10.1186/s12968-017-0361-7

Figure Lengend Snippet: Injury Patterns on Histology. Images show TTC, Thioflavin S and H E stained short axis sections obtained from explanted hearts at 24 h post intervention. TTC sections indicate region on necrosis appearing white in the I-HEM group (no hemorrhage) and reddish white in I+HEM group (hemorrhage); the +HEM group did no show infarction. Under ultra-violet light, non-fluorescent regions on the Thioflavin S stain highlight areas of compromised endothelium as seen in the +HEM and I+HEM groups; this confirms presence of microvascular damage in the hemorrhage groups. The H E images were obtained from the region of interest shown on the TTC stained sections. Hemorrhage was apparent in the +HEM and I+HEM groups as evidenced from the interstitial distribution of red blood cells ( open arrow heads ) in the affected LAD territory; red blood cells were absent in the I-HEM group. Edematous development was observed in all three groups ( arrows ) along with the presence of inflammatory cells ( closed arrow heads )

Article Snippet: Prior to sacrifice, a 4% solution of fluorescent dye thioflavin S (1 ml/kg, Sigma-Aldrich) was intravenously injected as a bolus to examine endothelial cell distribution and arterial patency in the heart [ ].

Techniques: Staining

Journal: Endocrinology

Article Title: Accumulation of Intrahepatic Islet Amyloid in a Nonhuman Primate Transplant Model

doi: 10.1210/en.2011-1560

Figure Lengend Snippet: Native pancreas biopsies stained for insulin ( red ) and thioflavin S ( green ) are demonstrated from normal monkey (A), normal human (B), and type 2 diabetic human (C). Native pancreas biopsies stained for insulin ( red ) and IAPP ( green ) are also demonstrated from normal monkey (D), normal human (E), and type 2 diabetic human (F), where intracellular colocalization of insulin and IAPP appears orange . Only the type 2 diabetic pancreas with amyloid present by thioflavin S evidences additional IAPP present extracellularly immediately surrounding β-cells. Intrahepatic islet biopsies stained for insulin ( red ) and IAPP ( green ) similarly show intracellular colocalization of insulin and IAPP in a biopsy without amyloid (G), and intra- and extracellular IAPP in a biopsy with amyloid (H). In I, separate images for insulin ( red , 1), IAPP ( green , 2), and their combination (3) are shown.

Article Snippet: Islet amyloid was detected by staining with thioflavin S (5 g/1000 ml solution; Sigma-Aldrich, St. Louis, MO) as previously described ( ) using normal cynomolgus and human pancreata as negative controls and type 2 diabetic human pancreata as positive controls ( ).

Techniques: Staining

Journal: Endocrinology

Article Title: Accumulation of Intrahepatic Islet Amyloid in a Nonhuman Primate Transplant Model

doi: 10.1210/en.2011-1560

Figure Lengend Snippet: Intrahepatic islet transplant biopsies stained for insulin ( red ) and thioflavin S ( green ) are shown from recipient T4536 at posttransplant months 18 (A), 40 (B), and 46 (C), recipient 211050 at month 4 (D), recipient 93–100 at month 10 (E), recipient 3845 at month 43 (F), and recipient CR14 at months 14 (G), 48 (H), and 66 (I). Apparent is an increased accumulation of islet amyloid and decrease in islet β-cells when examining the same animals (A–C and G–I) or different animals (D–F) over time. Magnification, ×40. mo, Month.

Article Snippet: Islet amyloid was detected by staining with thioflavin S (5 g/1000 ml solution; Sigma-Aldrich, St. Louis, MO) as previously described ( ) using normal cynomolgus and human pancreata as negative controls and type 2 diabetic human pancreata as positive controls ( ).

Techniques: Staining

Journal: Endocrinology

Article Title: Accumulation of Intrahepatic Islet Amyloid in a Nonhuman Primate Transplant Model

doi: 10.1210/en.2011-1560

Figure Lengend Snippet: Images shown are multilayer composites created by recoloring and scaling spectrally unmixed label component planes. A, Mask overlay indicates where inForm pattern-recognition software detected islet tissue. B, Insulin segmentation mask indicates the detection of islet β-cells as green fill. C, Thioflavin S segmentation mask indicates the detection of islet amyloid as green fill, with the islet representing an intrahepatic islet affected with amyloid from monkey CR14 at 66 months after transplant. The corresponding panels D–F demonstrate the software-detected islet tissue, β-cells, and amyloid, respectively, from a normal human islet not affected with amyloid.

Article Snippet: Islet amyloid was detected by staining with thioflavin S (5 g/1000 ml solution; Sigma-Aldrich, St. Louis, MO) as previously described ( ) using normal cynomolgus and human pancreata as negative controls and type 2 diabetic human pancreata as positive controls ( ).

Techniques: Software