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Immunofluorescence
Published in Guy Cox, Fundamentals of Fluorescence Imaging, 2019
There are several methods for preparing cell cultures for immunofluorescences. Cell cultures tend to be divided into adherent and suspension cultures and this will affect how cultures are processed for immunofluorescence. Cell cultures can be stained directly or dissociated cells can be spun directly onto slides using a cytocentrifuge before immunostaining. Cell cultures can also be concentrated using centrifugation and cell blocks prepared either for cryotomy or after formalin fixation, processed to paraffin for routine microtomy and immunostaining [29].
Efficacy of polyvinylpyrrolidone-capped gold nanorods against 7,12 dimethylbenz(a)anthracene-induced oviduct and endometrial cancers in albino rats
Published in Egyptian Journal of Basic and Applied Sciences, 2023
Hend Gamal, Walid Tawfik, Hassan H El-Sayyad, Heba Mohamed Fahmy, Ahmed N. Emam, Heba A El-Ghaweet
In the oviduct, Ki-67 immunohistochemistry revealed higher immunostaining in the DMBA carcinogenesis group, indicating more unusual proliferative activity (Figure 10d,e). However, when the DMBA carcinogenesis group was treated with PVP-capped AuNRs, the Ki-67 immunohistochemical reactivity was reduced (Figure 10f). Following image analysis, Ki-67 showed that the immune response to DMBA carcinogenesis was significantly higher than in the other groups that had been studied (Figure 8). Also, the uterine endometrium of negative control possessed weak-to-moderate positive reaction for Ki-67 (Figure 11a) compared to more reaction post-DMBA carcinogenesis injection (Figure 11c,d). In contrast, improved weak reaction was remarked in DMBA carcinogenesis treated with PVP-capped AuNRs (Figure 11e,f). Ki-67 displayed an essential increase in the immune reaction in the DMBA carcinogenesis, improved in DMBA carcinogenesis treated with PVP-capped AuNRs group but still did not match with control values (Figure 12).
Effects of whole-body vibration on reproductive physiology in a rat model of whole-body vibration
Published in Journal of Toxicology and Environmental Health, Part A, 2022
K. Krajnak, S. Waugh, D. Welcome, X.S. Xu, C. Warren, W. McKinney, R.G. Dong
Oxidative stress: Concentrations of reactive oxygen species (ROS) were measured in the pituitary, ovaries, testes, uteri, and prostates; (Table 1). Because one ovary was used for PCR and the other for histology, oxidative stress in the ovary was estimated by the intensity of nitrotyrosine immunostaining in sections. In all other tissues, ROS concentrations were measured using the fluorescent dye, 2ʹ7’-dichlorofluorescien diacetate (DCFDA; (Krajnak et al. 2020)). In females, exposure to WBV did not markedly affect nitrotyrosine labeling in ovaries or fluorescence in the pituitary. However, exposure to WBV resulted in an increase in ROS (fluorescence) in uteri. In males, exposure to WBV did not alter ROS in the pituitary or testes but produced significant reduction in ROS-induced fluorescence in the prostate.
Decellularization and genipin crosslinking of amniotic membrane suitable for tissue engineering applications
Published in Journal of Biomaterials Science, Polymer Edition, 2018
Sarumathi Gobinathan, Siti Solehah Zainol, Siti Fatmah Azizi, Nabil Mohamad Iman, Rajasegaran Muniandy, Hanis Nazihah Hasmad, Mohd Reusmaazran bin Yusof, Salina Husain, Haslinda Abd Aziz, Yogeswaran Lokanathan
This study showed that the decellularization of AM using thermolysin and NaOH did not lead to an apparent decrease in the extracellular matrix (ECM), especially collagen content of the AM. Collagen is an important component for structural support, cell anchorage, cell proliferation and tissue formation [31, 32]. HAM is found to contain Collagen type I, III, IV, V and VI [4]. Thus, preserving the ECM content of AM is vital in development of scaffold for tissue engineering applications and the genipin crosslinking of the dAM may further enhance the preservation of the ECM on the decellularized membrane [29]. It is also assumed that this crosslinking process enable preservation of the other native growth factors as previous studies have shown that genipin crosslinking preserved various ECM components and growth factors on the decellularized porcine liver tissue [29]. In this study, we did not perform the immunostaining for collagen on genipin-crosslinked AM due to the auto-fluorescence of the genipin crosslinked tissues. Previous study conducted by Hwang et al. [33], showed that incubation with genipin induced strong auto-fluorescence within collagen hydrogels [33].