Novel fluorescent staining protocol for thick sections of human osteochondral tissues to facilitate correlation with MRI and CT

2021 ◽  
Author(s):  
Qingbo Tang ◽  
Alecio F. Lombardi ◽  
Nicole Le ◽  
Jonathan H. Wong ◽  
Judith L. Williams ◽  
...  
Keyword(s):  
Fluorescent Staining ◽  
Staining Protocol

Fading of the diaminobenzidine reaction product in the confocal scanning laser microscope

1989 ◽  
Vol 47 ◽  
pp. 146-147
Author(s):  
JS Deitch ◽  
KL Smith ◽  
JW Swann ◽  
JN Turner
Keyword(s):  
Pyramidal Neurons ◽  
Light And Electron Microscopy ◽  
Scanning Laser ◽  
Laser Exposure ◽  
Confocal Scanning Laser Microscope ◽  
Fluorescent Markers ◽  
Before And After ◽  
Staining Protocol ◽  
Confocal Scanning ◽  
Confocal Scanning Laser

Neurons labeled with horseradish peroxidase and reacted with diaminobenzidine (DAB) can be imaged using a confocal scanning laser microscope (CSLM) in the reflection mode. In contrast to fluorescent markers, the DAB reaction product is thought to be stable and can be observed by both light and electron microscopy. We have investigated the sensitivity of the DAB reaction product to laser irradiation, and present the spectrophotometric properties of DAB before and after exposure in the CSLM.Pyramidal neurons in slices of rat hippocampus were injected with biocytin (a biotin-lysine complex), fixed overnight in 4% paraformaldehyde, and vibratome sectioned at 75 μm. Biocytin was detected with avidin-HRP (1:200) in 0.5% Triton X-100, incubated in DAB (0.5 mg/ml) with or without 0.04% nickel ammonium sulfate (Ni), dehydrated, and imaged in a Bio Rad MRC-500 CSLM with an argon ion laser (488 and 514 nm). Spectrophotometric measurements of the soma were made on a Zeiss microspectrophotometer, as a function of laser exposure (100-1000 scans) and staining protocol.

scholarly journals Novel approach for quantification of multiple immunofluorescent signals using histograms and 2D plot profiling of whole-section panoramic images

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Roko Duplancic ◽  
Darko Kero
Keyword(s):  
Histological Section ◽  
Cell Counting ◽  
Tissue Samples ◽  
Spatial Gradients ◽  
Multiple Markers ◽  
Novel Approach ◽  
Academic Settings ◽  
Panoramic Images ◽  
Multiple Regions ◽  
Staining Protocol

AbstractWe describe a novel approach for quantification and colocalization of immunofluorescence (IF) signals of multiple markers on high-resolution panoramic images of serial histological sections utilizing standard staining techniques and readily available software for image processing and analysis. Human gingiva samples stained with primary antibodies against the common leukocyte antigen CD45 and factors related to heparan sulfate glycosaminoglycans (HS GAG) were used. Expression domains and spatial gradients of IF signals were quantified by histograms and 2D plot profiles, respectively. The importance of histomorphometric profiling of tissue samples and IF signal thresholding is elaborated. This approach to quantification of IF staining utilizes pixel (px) counts and comparison of px grey value (GV) or luminance. No cell counting is applied either to determine the cellular content of a given histological section nor the number of cells positive to the primary antibody of interest. There is no selection of multiple Regions-Of-Interest (ROIs) since the entire histological section is quantified. Although the standard IF staining protocol is applied, the data output enables colocalization of multiple markers (up to 30) from a given histological sample. This can serve as an alternative for colocalization of IF staining of multiple primary antibodies based on repeating cycles of staining of the same histological section since those techniques require non standard staining protocols and sophisticated equipment that can be out of reach for small laboratories in academic settings. Combined with the data from ontological bases, this approach to quantification of IF enables creation of in silico virtual disease models.

scholarly journals Suicide by intoxication with iron (III) chloride

2021 ◽  
Author(s):  
Sławomir Majdanik ◽  
Barbara Potocka-Banaś ◽  
Sebastian Glowinski ◽  
Krzysztof Borowiak
Keyword(s):  
Prussian Blue ◽  
Chloride Ions ◽  
Iron Chloride ◽  
Histopathological Analysis ◽  
Fatal Intoxication ◽  
Internal Organs ◽  
Specific Staining ◽  
Iron Supplements ◽  
Staining Protocol ◽  
Hematoxylin And Eosin

Abstract Purpose Cases of iron intoxication are not very often encountered in toxicology practice, and most of those reported concern accidental intoxications with iron supplements in young children. The paper presents a rare case of a suicide by intoxication in an adult woman who ingested a solution of iron (III) chloride. Methods A forensic was at the Department of Forensic Medicine, PMU in Szczecin. Toxicology tests of blood sampled from the deceased were performed using a 644 CIBA CORNING ion selective analyzer and proprietary reagent kits. Histopathological was with the use of the standard staining protocol (hematoxylin and eosin) and staining specific for iron (Prussian blue). Results Autopsy revealed a distinct yellow discolouration and thrombotic necrosis of the oral mucosa and almost the whole gastrointestinal tract, as well as similar changes in the adjacent internal organs. Considerably high levels of iron and chloride ions were detected in specimens of internal organs preserved during autopsy. Histopathological analysis performed with the use of staining specific for iron (Prussian blue) also confirmed the presence of iron in the examined tissues, especially in the intestines and liver. Conclusions Considering the above findings, it was concluded in the forensic report that the death of the woman was caused by the ingestion of iron chloride. The reported case of fatal intoxication is one of the few described in the literature, and its course implies that in the case of initially diagnosed intoxication with corrosive compounds, the possibility of using metal-containing poison should also be considered in the differential diagnosis. In addition to routine toxicological tests performed in fatal cases we also draw attention to the possibility of using specific staining protocols for microscopic specimens.

Annual Cycle of Freshwater Diatoms in the High Arctic Revealed by Multiparameter Fluorescent Staining

2020 ◽  
Vol 80 (3) ◽  
pp. 559-572
Author(s):  
Eva Hejduková ◽  
Josef Elster ◽  
Linda Nedbalová
Keyword(s):  
Annual Cycle ◽  
High Arctic ◽  
Fluorescent Staining ◽  
Freshwater Diatoms

scholarly journals Comparison of the Benzanthrone Luminophores: They Are Not Equal for Rapid Examination of Parafasciolopsis fasciolaemorpha (Trematoda: Digenea)

Biomolecules ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 598
Author(s):  
Ilze Rubenina ◽  
Inese Gavarane ◽  
Elena Kirilova ◽  
Ligita Mezaraupe ◽  
Muza Kirjusina
Keyword(s):  
Tertiary Amine ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Amine Group ◽  
Tertiary Amine Group ◽  
Organ Systems ◽  
Staining Protocol ◽  
Anatomical Arrangement ◽  
Moose Population

Luminescent derivatives of benzanthrone are becoming more useful based on their light-absorbing and fluorescent-emitting properties. Our previous studies showed that luminescent staining properties of the same benzanthrone dye differ for variable parasite samples. Therefore, two types of benzanthrone dyes were prepared. One has a strongly basic amidine group and a halogen atom, and the other has an amide moiety and a tertiary amine group. Trematoda Parafasciolopsis fasciolaemorpha is a liver fluke of a moose (Alces alces) and has a significant influence on the health and abundance of the moose population. Staining protocols for parasite P. fasciolaemorpha specific organ or organ systems imaging are mostly time-consuming and labor-intensive. The study aimed to compare the fixation technique and the staining protocol by synthesized benzanthrone luminescent dyes to determine detailed morphology, anatomical arrangement of the organ systems and gross organization of the muscle layers of P. fasciolaemorpha using confocal laser scanning microscopy. Luminophores were tested for samples fixed in different fixatives. Developed dyes and staining protocol resulting in imaging of all parts of trematode without additional sample preparation procedures, which usually are required for parasite examination. Obtained results confirmed that the most qualitative results could be reached using 3-N-(2-piperidinylacetamido)benzanthrone dye which has amide moiety and a tertiary amine group. Based on obtained results, 3-N-(2-piperidinylacetamido)benzanthrone gave more qualitative parasite visualization than 2-bromo-3-N-(N′,N′-dimethylformamidino)benzanthrone.

Development of a simple and effective silver staining protocol for detection of DNA fragments

2017 ◽  
Vol 38 (8) ◽  
pp. 1175-1178 ◽  
Author(s):  
Wenjie Liu ◽  
Ronghua Li ◽  
Habtamu Ayalew ◽  
Yanshi Xia ◽  
Guihua Bai ◽  
...  
Keyword(s):  
Silver Staining ◽  
Dna Fragments ◽  
Staining Protocol
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