China
Africa
Explore chapters and articles related to this topic
Methods for Evaluating Articular Cartilage Quality
Published in Kyriacos A. Athanasiou, Eric M. Darling, Grayson D. DuRaine, Jerry C. Hu, A. Hari Reddi, Articular Cartilage, 2017
Kyriacos A. Athanasiou, Eric M. Darling, Grayson D. DuRaine, Jerry C. Hu, A. Hari Reddi
Dyes commonly used for staining cartilage glycosaminoglycans take advantage of the negative charge of molecules, and these include Safranin O, toluidine blue, and Alcian blue (Figure 5.17). Safranin O is a red to orange stain, often used with Fast Green (a neutral dye) as a counterstain. Articular cartilage should first be protonated in an acidic solution before dye binding occurs. Toluidine blue is a metachromatic dye that changes from blue to pink-purple when the sulfated groups of a glycosaminoglycan molecule bring several dye molecules close together. This color change is only visible under aqueous conditions, and dried toluidine blue complexes will appear blue. Toluidine blue has been used in cartilage histology with aqueous mounts to distinguish sulfated glycosaminoglycans from other anionic species.
Cytological and Histological Analysis by Image Processing
Published in Corain Livio, Arboretti Rosa, Bonnini Stefano, Ranking of Multivariate Populations, 2017
Corain Livio, Arboretti Rosa, Bonnini Stefano
Skin samples 10 cm2 wide and 1 cm thick were collected from the hips of the animals using a scalpel blade, and fixed for 24 h in 4% formaldehyde. Each sample was reduced to fragments of 1 cm2, dehydrated and embedded in paraffin. From each sample, transverse sections of 4 μm thickness were obtained using a Leica R M2035 microtome. Slides were stained with conventional laboratory methods: (1) haematoxylin and eosin (H&E) for identification of general structure; (2) Masson’s trichrome technique and (3) orcein staining, to evaluate degeneration of collagen and elastic fibers and (4) periodic acid-Schiff-positive technique (Alcian Blue - PAS) to detect neutral and acidic polysaccharides. All methods were performed according to Bonucci (1981). The sections were then dehydrated and cover-slipped with balsam for light microscopy, and subsequently visualized using both 10 × and 20 × optical zooms with an Olympus DP70 RGB camera controlled by Cell Software (Olympus), interfacing a microscope Olympus Vanox AHBT3. Figure 8.6 displays several sample images by stained methods, representing slides of fresh (TO), refrigerated (Tl), short time salted (T2) and long time salted hides (T3).
Methods for Analyzing Floc Properties
Published in Ian G. Droppo, Gary G. Leppard, Steven N. Liss, Timothy G. Milligan, FLOCCULATION in NATURAL and ENGINEERED ENVIRONMENTAL SYSTEMS, 2004
Ian G. Droppo, Gary G. Leppard, Steven N. Liss, Timothy G. Milligan
Arange of stains with specificity for beta-D-glucan polysaccharides are used as general stains, these include calcofluor white and congo red. Ruthenium red has also been used as a light microscopy stain for detection of EPS. Probes for glycoaminoglycan such as Alcian blue may also be used as a general stain for “polysaccharides.” Wetzel et al. 35 demonstrated its use for determination of total EPS in microbial biofilms, in this case it was used indirectly and not for microscopy. Due to the complexity of the EPS the likelihood of finding a true total polysaccharide probe appears to be very limited. “L1615_C006” - 2004/11/19- 18:49- page 127-#7
Three-dimensional duck’s feet collagen/PLGA scaffold for chondrification: role of pore size and porosity
Published in Journal of Biomaterials Science, Polymer Edition, 2018
Jeong Eun Song, Nirmalya Tripathy, Se Rom Cha, Sung Hyun Jeon, Soon Yong Kwon, Dong Sam Suh, Gilson Khang
In order to confirm the proliferation of costal chondrocytes in the 80 wt% DC/PLGA scaffolds according to the pore size in in vivo environment, 80 wt% DC/PLGA scaffolds at 1 and 4 weeks after implant was extracted and H & E, Alcian Blue, and Safanin-O staining were performed, as shown in Figure 4. A circular lacuna, a marker of chondrocytes, was observed on all 80 wt% DC/PLGA scaffolds at 7 days. High numbers of costal chondrocytes were formed in aggregates in 80 wt% DC/PLGA scaffolds with 250–355 and 355–425 μm pore sizes as compared to the 80 wt% DC/PLGA scaffolds with 90–180 and 180–250 μm pore sizes. At 28th day, costal chondrocytes were uniformly distributed on all 80 wt% DC/PLGA scaffolds, and the number of costal chondrocyte was significantly increased than 7th day. In addition, it was confirmed that the tissue was regenerated between the pore as the pore size increased. The alcian blue staining implies the glycosaminoglycan (GAG) expression, which is an acidic mucous polysaccharide secreted by chondrocytes. It was confirmed that the blue stained lacuna of chondrocytes at 7th days are clearly distributed in some places in 80 wt% DC/PLGA scaffolds. In addition, the amount of chondrocytes as well as the GAG expressions was increased at 28 days. Safranin-O staining is also performed to confirm GAG expression. Similar to alcian blue staining, it was confirmed that the chondrocytes were reddish in places, and 28 days showed a wider range than 7 days. In particular, the coloring is better in 80 wt% DC/PLGA scaffolds with 250–355 and 355–425 μm pore sizes, the chondrocytes seeded in a large porous size 80 wt% DC/PLGA scaffolds were proliferated more and amount of GAG secretion was increased, forming an ECM and inducing tissue organization of the 80 wt% DC/PLGA scaffolds
A human pericardium biopolymeric scaffold for autologous heart valve tissue engineering: cellular and extracellular matrix structure and biomechanical properties in comparison with a normal aortic heart valve
Published in Journal of Biomaterials Science, Polymer Edition, 2018
Frantisek Straka, David Schornik, Jaroslav Masin, Elena Filova, Tomas Mirejovsky, Zuzana Burdikova, Zdenek Svindrych, Hynek Chlup, Lukas Horny, Matej Daniel, Jiri Machac, Jelena Skibová, Jan Pirk, Lucie Bacakova
For the histological evaluation, the samples were fixed in 10% formalin, dehydratated and embedded in paraffin wax, cut at 4 μm and stained with Hematoxylin-eosin (H&E) and Elastica Weigert – van Gieson. Alcian blue staining was used for staining acidic glycosaminoglycans (GAGs; mucopolysaccharides).