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Beta and Alpha Particle Autoradiography
Published in Michael Ljungberg, Handbook of Nuclear Medicine and Molecular Imaging for Physicists, 2022
Anders Örbom, Brian W. Miller, Tom Bäck
In 2017, Asano and colleagues investigated the distribution of selective glucocorticoid receptor agonist (SEGRA) SA22465 in the eyes and eyelids of rabbits, which were sectioned and mounted on slides pre-coated with photographic emulsion [31]. Before sacrifice, the rabbits were given 14C-SA22465 ophthalmic suspension into each eye. The slides containing 5 µm thick sections were exposed for 28 days and then developed and counter-stained with haematoxylin and eosin. The dark grains from autoradiography on top of the haematoxylin of different parts of the eye and eyelid can be seen in Figure 30.2. Emulsion autoradiography was also used by Aguero and colleagues on brain sections from deceased Alzheimer patients [32]. The sections were incubated with either the PET tracer 18F-MK-6240 or 18F-AV-1451 before the slides were covered with emulsion, one of two autoradiography methods employed, and exposed and developed.
Inflammation and Cytokines in Airway Wall Remodelling
Published in Alastair G. Stewart, AIRWAY WALL REMODELLING in ASTHMA, 2020
Endobronchial biopsy in asthma has the distinct advantages of accurate characterisation of asthmatic subjects and the possibility of repeated intervention. Early studies involved the more invasive procedure of rigid bronchoscopy.25,26 However, fibreoptic bronchoscopy has made endobronchial biopsy a relatively safe and tolerable procedure.22,23 Limitations of this technique include epithelial sampling without submucosal tissue, epithelial damage, and crush artefact. Identification of specific cell types is usually unreliable in standard haematoxylin and eosin-stained sections (Figure 1). Specific monoclonal antibodies enable the accurate enumeration of cells (Figures 2 and 3).
Laboratory Procedures and Management
Published in Jeremy R. Jass, Understanding Pathology, 2020
The early attempts at tissue staining were achieved by trial and error using natural dyes that had been available and in use for centuries, if not millennia, for dying fabrics. Leeuwenhoek (1632–1723) applied saffron solution to preparations of muscle fibres. By the end of the nineteenth century, the most popular stain for tissue sections was carmine derived from cochineal (Mayer, 1892). Cochineal is a red dye prepared from the dried female bodies of a scale insect, Dactylopius coccus. It was known to the Aztecs, the ancient Romans and apparently in biblical times since the Divinity exhorted Moses to prepare offerings of rams’ skins dyed red (Exodus 25:5). Orcein, known originally as French purple, dates from the 1300s (AD) when it was prepared from an extract of lichen (a primitive plant that is part fungus and part alga) that was exposed to air in the presence of ammonia formed in fermented urine (Conn, 1948). Orcein is still used for staining various tissue components, but thankfully is now prepared differently. Haematoxylin is derived from the wood of a tree called Haematoxylon campechianum, so named because it originated in the Mexican State of Campeche. Synthetic dyes, for example alcian blue developed by ICI, have also been used to stain cell products.
Network pharmacology approach to investigate the multitarget mechanisms of Zhishi Rhubarb Soup on acute cerebral infarction
Published in Pharmaceutical Biology, 2022
Yuejia Shao, Yue Zhang, Rongrong Wu, Lurui Dou, Fengjiao Cao, Yuqing Yan, Yuming Tang, Chi Huang, Yang Zhao, Jinghua Zhang
Six rats from per group were used for this analysis. Sections of the infarcted brain were fixed with 4% paraformaldehyde at room temperature for 24 h and then embedded in paraffin. A series of 5 µm thick sections were cut from the block. Paraffin sections were deparaffinized and hydrated through a series of graded alcohol solutions. Tissue slices were then rinsed with PBS (0.1 M, pH 7.4, 3 × 5 min) and incubated at room temperature with BSA for 30 min. The closure solution was removed, and primary antibody (NF-κB p65 and RAGE) is added dropwise on the sections. Samples were placed flat in a wet box and incubated overnight at 4 °C. The samples were washed with PBS, and corresponding secondary antibodies were added in a dropwise fashion. The samples were incubated at room temperature for 50 min and washed, then DAB chromogenic solution was added. The sample was restained with haematoxylin for 3 min. The nuclei were stained blue with haematoxylin. Positive DAB expression appeared brownish-yellow, and the field of view was selected for image acquisition using a Nikon imaging system (Tokyo, Japan). The result was presented as the positive cell density: number of positive cells/tissue area to be measured.
Evaluation of different haematoxylin stain subtypes for the optimal microscopic interpretation of cutaneous malignancy in Mohs frozen section histological procedure
Published in British Journal of Biomedical Science, 2021
JA Gabriel, M Shams, GE Orchard
As a part of the Mohs procedure, H&E staining remains the staple method for microscopic evaluation for pathological diagnosis and interpretation of these tumour types. In most cases, the haematoxylin nuclear staining plays an essential role in determining neoplastic disease. The presence of basophilic, hyperchromatic nuclei, apoptotic bodies, mitotic figures and pleomorphism all rely on clear staining to allow the generation of unequivocal diagnoses. The haematoxylin dye is extracted from the bark of the logwood tree Haematoxylin Campechianum, originally located in the Mexican state Campeche [6]. The conversion of haematoxylin to haematin, vital for its ability to bind to nuclear components, is aided by the use of mordants. There are a broad range of mordants which can impact the tissue components stained and colour of staining, which is visualised. The mordants are usually a metal cation such as iron, aluminium, molybdenum, lead and tungsten [7].
In vivo percutaneous permeation of gold nanomaterials in consumer cosmetics: implication in dermal safety assessment of consumer nanoproducts
Published in Nanotoxicology, 2021
Mingjing Cao, Bai Li, Mengyu Guo, Ying Liu, Lili Zhang, Yaling Wang, Bin Hu, Jiayang Li, Duncan S. Sutherland, Liming Wang, Chunying Chen
HE staining is a popular and established method in histology. Herein it was used to evaluate the dermal lesions caused by exposure of cosmetic creams and extracted Au nanosheets. 4 μm serial paraffin sections sliced carefully from skin surface to subcutaneous layer were deparaffinized, rehydrated, and stained with hematoxylin and eosin. For IHC analysis, 4 μm serial sections were firstly deparaffinized and rehydrated, followed by retrieving the masked antigens with 0.01 M sodium citrate buffer in the microwave (96 °C for 10 min). After blocking endogenous peroxidase with 30% H2O2, the sections were incubated with primary antibodies overnight at 4 °C and followed by the incubation with secondary antibodies labeled by horseradish peroxidase (HRP) at 37 °C for 1 h. Hematoxylin was used for nuclei staining.