Beta and Alpha Particle Autoradiography
Michael Ljungberg in Handbook of Nuclear Medicine and Molecular Imaging for Physicists, 2022
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.
Laboratory Procedures and Management
Jeremy R. Jass in 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.
The ‘awe-full’ fascination of pathology
Alan Bleakley in Routledge handbook of the medical humanities, 2019
Working so close to death, in the living person and in the deceased body, we practise a wonderful, if not an ‘awe-full,’ medical specialty. How can we account for such a medical, scientific, and profoundly human enterprise that is the practice of pathology? It seems appropriate to us to respond in poetic voice (see Box below, ‘The awe-full fascination of pathology’). To situate this practice, it is worth noting that pathology arose as a discipline in the morgue. We question the corpse, the nature of the disease, reconstructing the course of events from the very early inception of the disease until death. As pathologists entered the world of microscopy, they characterised the cell as the unit of life. How ‘awesome/awe-full’ it must have been to look at the first slides on a microscope! How to make sense of that myriad of findings, the complex multitude of different cells, awaiting discovery and understanding? Today, we look at slide specimens routinely stained in haematoxylin and eosin. But in those early days, multiple different dyes would have been tried and the pathology researcher likely had an aesthetic sensibility to choose one for a routine use. Beyond routine stains, an array of special stains has been developed with specific purposes.
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.
Areca nut procyanidins prevent ultraviolet light B-induced photoaging via suppression of cyclooxygenase-2 and matrix metalloproteinases in mouse skin
Published in Drug and Chemical Toxicology, 2022
Chia-Ling Weng, Chih-Chiang Chen, Han-Hsing Tsou, Tsung-Yun Liu, Hsiang-Tsui Wang
To evaluate the effect of ANP on UVB-induced alterations in skin morphology, mice were sacrificed 24 h after the last UV exposure. Skin biopsies were obtained from the dorsal skin, perpendicular to the long axis of the trunk, fixed in 4% paraformaldehyde and processed for hematoxylin & eosin staining and microscopic evaluation. Hematoxylin stained for nuclei (dark blue) and eosin stained for cytoplasm (pink). For histopathological analyses of collagen fibers, skin samples were processed and stained with Masson’s trichrome for quantification of type I collagen. Briefly, skin samples were fixed in 4% paraformaldehyde overnight at room temperature and stained for collagen fibers using a trichrome stain (Masson) kit (Sigma, St. Louis, MO, USA) according to manufacturer’s instructions. Collagen was stained blue, nuclei were stained dark red, and cytoplasm or muscle was stained red. Quantification of collagen fibers in Masson Trichrome stained sections was measured using NIH Image J software, and expressed as the percentage area occupied by each fiber in the upper dermis.
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].
Related Knowledge Centers
- Cell Biology
- Chemical Compound
- Counterstain
- Histology
- Eosin
- Staining
- H&E Stain
- Papanicolaou Stain
- Redox
- Hematein