Enhancement of the periodic acid — Schiff (PAS) and periodic acid — thiocarbohydrazide — silver proteinate (PA-TCH-SP) reaction in LR White sections

Neiss, Wolfram F.

doi:10.1007/bf00570331

Cited by 15 publications

(4 citation statements)

References 22 publications

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“…Standard incubations were performed according to Thiéry as described . In case of weak staining (eg, Figure E), incubation with PA and thiocarbohydrazide (TCH) was optionally extended up to 15 hours each, and/or silver proteinate treatment (SP) was carried out at +50°C . Alternatively, formaldehyde‐fixed biopsies were subjected to high‐pressure freezing, freeze‐substitution and resin embedding as described for native cell samples, followed by standard incubations (60 minutes PA, 120 minutes TCH, 30 minutes SP at RT: Reference )—an approach, that yielded considerably intensified PAS‐reaction product (Figure F).…”

Section: Methodsmentioning

confidence: 99%

Abnormal Rab11‐Rab8‐vesicles cluster in enterocytes of patients with microvillus inclusion disease

Vogel

Janecke

Krainer

et al. 2017

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Microvillus Inclusion Disease (MVID) is a congenital enteropathy characterized by accumulation of vesiculo-tubular endomembranes in the subapical cytoplasm of enterocytes, historically termed “secretory granules”. However, neither their identity nor pathophysiological significance is well defined. Using immunoelectron microscopy and tomography we studied biopsies from MVID patients (3x Myosin 5b mutations, 1x Syntaxin3 mutation) and compared them to controls and genome-edited CaCo2 cell models, harboring relevant mutations. Duodenal biopsies from two patients with novel Myosin 5b mutations and typical clinical symptoms showed unusual ultrastructural phenotypes: aberrant subapical vesicles and tubules were prominent in the enterocytes, though other histological hallmarks of MVID were almost absent (ectopic intra-/intercellular microvilli, brush border atrophy). We identified these enigmatic vesiculo-tubular organelles as Rab11-Rab8-positive recycling compartments of altered size, shape and location harboring the apical SNARE Syntaxin3, apical transporters Sodium-Hydrogen Exchanger 3 (NHE3) and cystic fibrosis transmembrane conductance regulator (CFTR). Our data strongly indicate that in MVID disrupted trafficking between cargo vesicles and the apical plasma membrane is the primary cause of a defect of epithelial polarity and subsequent facultative loss of brush border integrity, leading to malabsorption. Furthermore, they support the notion that mislocalization of transporters, such as NHE3 substantially contributes to the reported sodium loss diarrhea.

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Section: Methodsmentioning

confidence: 99%

Abnormal Rab11‐Rab8‐vesicles cluster in enterocytes of patients with microvillus inclusion disease

Vogel

Janecke

Krainer

et al. 2017

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“…Note that SP-treatment at RT worked only with quite old SP batches (>30-year-old). Actual batches require heating not to fail, i.e., incubation at +50 °C for 30 min [ 50 ] yielded good results.…”

Section: Methodsmentioning

confidence: 99%

Advanced Microscopy for Liver and Gut Ultrastructural Pathology in Patients with MVID and PFIC Caused by MYO5B Mutations

Hess

Krainer

Filipek

et al. 2021

JCM

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Mutations in the actin motor protein myosinVb (myo5b) cause aberrant apical cargo transport and the congenital enteropathy microvillus inclusion disease (MVID). Recently, missense mutations in myo5b were also associated with progressive familial intrahepatic cholestasis (MYO5B-PFIC). Here, we thoroughly characterized the ultrastructural and immuno-cytochemical phenotype of hepatocytes and duodenal enterocytes from a unique case of an adult MYO5B-PFIC patient who showed constant hepatopathy but only periodic enteric symptoms. Selected data from two other patients supported the findings. Advanced methods such as cryo-fixation, freeze-substitution, immuno-gold labeling, electron tomography and immuno-fluorescence microscopy complemented the standard procedures. Liver biopsies showed mislocalization of Rab11 and bile canalicular membrane proteins. Rab11-positive vesicles clustered around bile canaliculi and resembled subapical clusters of aberrant recycling endosomes in enterocytes from MVID patients. The adult patient studied in detail showed a severe, MVID-specific enterocyte phenotype, despite only a mild clinical intestinal presentation. This included mislocalization of numerous proteins essential for apical cargo transport and morphological alterations. We characterized the heterogeneous population of large catabolic organelles regarding their complex ultrastructure and differential distribution of autophagic and lysosomal marker proteins. Finally, we generated duodenal organoids/enteroids from biopsies that recapitulated all MVID hallmarks, demonstrating the potential of this disease model for personalized medicine.

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“…Four pieces (5 ϫ 1 mm) were routinely processed for light microscopy and/or transmission electron microscopy by postfixation in uranyl acetate, partial dehydration through graded ethanol mixtures to 70%, infiltration with hard-grade LR White acrylic resin (Light Resin Co., Reading, UK), embedding in acrylic resin, and cold catalytic polymerization at 4°C (20). Semithin (0.35-m) sections for light microscopy were stained with 0.5% toluidine blue for morphologic assessments and were stained by using a modification of the periodic acid thiocarbohydrazide-silver proteinate-silver enhancement (PATCH-SP-SE) method to facilitate observation of small blood vessels and capillaries (21).…”

Section: Biopsy Collection and Processingmentioning

confidence: 99%

Morphologic Changes in the Peritoneal Membrane of Patients with Renal Disease

Williams

Craig

Topley

et al. 2002

Journal of the American Society of Nephrology

861

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ABSTRACT. This study examined the morphologic features of the parietal peritoneal membranes of 130 patients undergoing peritoneal dialysis (PD) and compared them with the features of the peritoneal membranes of normal individuals, uremic predialysis patients, and patients undergoing hemodialysis. The median thickness of the submesothelial compact collagenous zone was 50 μm for normal subjects, 140 μm for uremic patients, 150 μm for patients undergoing hemodialysis, and 270 μm for patients undergoing PD (P < 0.001 for all versus normal subjects). Compact zone thickness increased significantly with the duration of PD therapy [0 to 24 mo, 180 μm (n = 58); 25 to 48 mo, 240 μm (n = 24); 49 to 72 mo, 300 μm (n = 13); 73 to 96 mo, 750 μm (n = 16); >97 mo, 700 μm (n = 19)]. Vascular changes included progressive subendothelial hyalinization, with luminal narrowing or obliteration. These changes were absent in samples from normal subjects but were present in 28% of samples from uremic patients and 56% of biopsies from patients undergoing PD. In the PD group, the prevalence of vasculopathy increased significantly with therapy duration (P = 0.0001). The density of blood vessels per unit length of peritoneum was significantly higher for patients with membrane failure and was correlated with the degree of fibrosis (P = 0.01). For the first time, a comprehensive cross-sectional analysis of the morphologic changes in the peritoneal membranes of patients undergoing PD is provided. The infrequency of fibrosis in the absence of vasculopathy suggests that vasculopathy may predispose patients to the development of fibrosis. This study provides a sufficiently large cohort of samples to allow structure-function relationships to be established, as well as providing a repository of tissue for further studies.

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Enhancement of the periodic acid — Schiff (PAS) and periodic acid — thiocarbohydrazide — silver proteinate (PA-TCH-SP) reaction in LR White sections

Cited by 15 publications

References 22 publications

Abnormal Rab11‐Rab8‐vesicles cluster in enterocytes of patients with microvillus inclusion disease

Abnormal Rab11‐Rab8‐vesicles cluster in enterocytes of patients with microvillus inclusion disease

Advanced Microscopy for Liver and Gut Ultrastructural Pathology in Patients with MVID and PFIC Caused by MYO5B Mutations

Morphologic Changes in the Peritoneal Membrane of Patients with Renal Disease

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