Methods for the Morphological Study of Tracheal and Bronchial Glands
Joan Gil in Models of Lung Disease, 2020
The presence of both neutral and acid glycoproteins has been demonstrated in the secretory cells of the submucosal glands. Of the various staining techniques available, the periodic acid-Schiff (PAS) and alcian blue (AB) techniques are much used; PAS is used to demonstrate the presence of neutral glycoprotein and AB for acid glycoprotein. Alcian blue, used at pH 2.6, stains both sialic acid and sulfate radicals bright blue. The control of AB staining, by varying the pH between 0.5 and 2.6, has been found by Jones and Reid (1973a, 1973b) to be satisfactorily selective, differentiating between sialo- and sulfomucin. With a combination of AB (pH 2.6) and PAS stains, mucous cells stain blue for acid glycoprotein. The mucous cells of the human tracheobronchial glands have been shown by Lamb and Reid (1972) to produce four groups of acid glycoprotein: sialomucin susceptible to sialidase; sialomucin resistent to sialidase; a sulfate identified by AB staining after acid hydrolysis (which removes all the sialomucin) but that is not stained by stains specific for sulfate; and a sulfate that stains after hydrolysis and stains with the specific stains for it. Ferret tracheal glands develop from intraepithelial cellular aggregates devoid of secretory granules at birth into complex, submucosal tubuloacinar structures composed predominantly of cells containing nonacidic (staining with PAS but not AB) secretory granules at 28 days (Leigh et al., 1986b), and thereafter acidic histochemical staining properties increase in secretory cells (Leigh et al., 1986a).
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.
Comparative Anatomy and Development of the Mammalian Disc
Peter Ghosh in The Biology of the Intervertebral Disc, 2019
The fine structure of the inner annulus shows that the inner annulus cells of the 2-month-old cat72 are either fibrocytes or chondrocytes, though some cells are degenerating. They contain intracytoplasmic filaments, microtubules, and occasional centrioles, as well as the usual organelles. The plasma membrane contains numerous micropinocytotic vesicles. In older animals, cellular organelles decrease and degenerate cells increase in number. At the same time, “bays” in the cell surface (Figure 9b) increase and become continuous with the dilated endoplasmic reticulum in degenerate cells (Figure 9c). Many cells are surrounded by granular densities (Figure 9b). These may be derived from the intracellular granular bodies72 and may represent the pericellular granules73 of light microscopy (Figure 9a). The fibrils that surround the cells sometimes show beading and banding in the cat72 as in other species.20,74 They are associated with material that is amorphous or finely granular in routine electron microscopic sections, but which stains densely with Alcian Blue62 (Figure 9d). Alcian Blue also stains a sheath around the collagen fibrils, a feature which indicates that glycosaminoglycans are associated closely with them.
Tensile Viscoelastic Properties of the Sclera after Glycosaminoglycan Depletion
Published in Current Eye Research, 2021
Hamed Hatami-Marbini, Mohammad Pachenari
The thickness of the strips in the control and No GAG group was 1.23 ± 0.13 mm and 1.25 ± 0.11 mm, respectively; these values are in agreement with those reported previously for porcine disks obtained from a similar region.29 The DMMB assay showed an about 88% decrease in GAG content because of the digestion process. The GAG content of samples in the control and No GAG groups was 6.39 ± 0.55 µg/mg and 0.79 ± 0.17 µg/mg, respectively. It is known that GAGs appears blue to bluish-green after staining with the Alcian blue. Figure 2a shows that GAGs existed throughout a typical scleral sample that was treated in buffer solution. However, Figure 2b shows that staining of Alcian blue was barely visible in a sample treated by the enzyme solution, i.e. the concentration of GAGs was significantly reduced in the samples treated with the enzyme.
A case report of a primary apocrine adenocarcinoma of the eyelid with literature review
Published in Orbit, 2018
C. Pagano Boza, R. Vigo, J. E. Premoli, J. Croxatto, J. Gonzalez Barlatay
Histopathology revealed a tumor with nodules of different sizes filled by enlarged cells, with pleomorphic and hyperchromatic nuclei, and eosinophillic cytoplasm. These cells were arranged forming glandular structures, abortive lights and cords, with mucoid secretion by decapitation, characteristic of these tumors. Scattered atypical mitosis was identified (Figure 2). Immunohistochemistry showed GCDFP-15 (protein 15 of the fibrocystic disease of the breast) positivity at the apical border of neoplastic cell (Figure 3a). CEA (carcinoembryonic antigen) and EMA (epithelial membrane antigen) were positive at the luminal cellular border (Figure 3b, c). Schiff’s Periodic Acid (PAS) and Alcian Blue were positive. These findings were consistent with Adenocarcinoma of Moll cells and free margins were obtained. Surprisingly, a new reconstructive procedure was not required because the left upper lid showed adequate function, cosmetic appearance and no signs of residual ptosis after the surgery (Figure 4). It was decided to perform inter-consultation with the Oncologist to do a screening. A computed tomography (CT) and positron emission tomography (PET) were performed and no evidenced of tumor elsewhere in the body was found. A close follow-up was carried out every month and 12 months later, the patient showed no evidence of recurrences.
Spatial analysis of gut microbiome reveals a distinct ecological niche associated with the mucus layer
Published in Gut Microbes, 2021
Kellyanne Duncan, Kelly Carey-Ewend, Shipra Vaishnava
7-μm thick slides were deparaffinized through a solution series for 5 minutes each (xylenes, xylenes, 100% EtOH, 95% EtOH, 75% EtOH, H2O). Alcian Blue/Pas staining was done using the protocol outlined in “Histotechnology: A Self-Instructional Text, 3rd edition. Pages 115–116”. Slides were stained with Alcian Blue for 30 minutes, washed with running tap water for 2 minutes followed by a rinse with deionized (DI) water. Slides were stained with 0.5% Periodic acid for 5 minutes, washed with DI, then stained with Schiff’s Reagent for 30 minutes followed by a rinse with running tap water for 3 minutes. Nuclei were stained with Hematoxylin for 2 minutes, washed with running water for 30 seconds, followed by 10 dips in Clarifier, another 30 second wash, 10 dips in Bluing Reagent, and a final 30 second wash. Slides went through a solution series for 5 minutes each (95% EtOH, 100% EtOH, Xylenes, Xylenes). Coverslips were applied with Cytoseal and slides were allowed to dry overnight in the fume hood. The dense mucus layer of 40 locations evenly distributed from available cross-sections on the slide (4–8 cross-sections) were measured for each mouse at 60X magnification to obtain the average mucus thickness for each mouse, then combined to obtain the overall average and standard deviation.
Related Knowledge Centers
- Cartilage
- Glycosaminoglycan
- Histology
- Sudan Stain
- Eosin
- Electron Microscope
- Staining
- Ion
- H&E Stain
- Periodic Acid–Schiff Stain