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Disease-Inspired Feature Design for Computer-Aided Diagnosis of Breast Cancer Digital Pathology Images
Published in de Azevedo-Marques Paulo Mazzoncini, Mencattini Arianna, Salmeri Marcello, Rangayyan Rangaraj M., Medical Image Analysis and Informatics: Computer-Aided Diagnosis and Therapy, 2018
To visualize these biomarkers, a DAB (3,3’-Diaminobenzidine) stain is used, which turns brown when the antibody of the stain binds to the antigen of the cell/tissue. Since tissue is essentially invisible under magnification, to visualize cells and tissue that do not have the biomarker(s), a counterstain is used. Once stained, pathologists use scoring systems to quantify the presence or absence of a particular biomarker.
Bioremediation of artificially contaminated soil with petroleum using animal waste: cow and poultry dung
Published in Cogent Engineering, 2020
O. Olawale, K. S. Obayomi, S. O. Dahunsi, O. Folarin
Gram’s Stain is one of the most frequently used processes in identifying bacteria and is used daily in hospitals. It is a primary test that quickly and cost-effectively divides bacteria into one of the two types: Gram-positive or Gram-negative. To identify the microbes grown, Gram staining technique was used to distinguish between the Gram’s positive and Gram’s negative organisms. A smear of bacteria was applied on to a slide, it was air-dried and then heated by passing it through a flame a few times. A few drops of crystal violet were added to the culture and allowed to stand for 1 min; the slide was rinsed in water for 5 sec. Gram’s iodine was then applied on the smear and allowed to stand for 1 min allowing the formation of dye-iodine complex in the cytoplasm of the bacteria cell. The slide was then tilted and decolorized with solvent (acetone solution) for 5 sec and then rinsed and shaken to remove excess. The slides were finally treated with safranin (counterstain since the safranin is lighter than crystal violet, it does not disrupt the purple coloration in Gram-positive cells) and allowed to stand for 1 min and then washed briefly with water and shaken off to remove excess. It was then allowed to dry before examining under a microscope.
Structural aspects of a trimetallic CuII derivative: cytotoxicity and anti-proliferative activity on human cancer cell lines
Published in Journal of Coordination Chemistry, 2019
Kuheli Das, Amitabha Datta, Chiara Massera, Catarina Roma-Rodrigues, Mariana Barroso, Pedro V. Baptista, Alexandra R. Fernandes
For autophagy analysis, 0.5 × 105 of HCT116 cells were seeded on top of a sterilized cover slide and let to adhere for 24 h. The supernatant was substituted with fresh medium containing 1 (at its IC50). For control purposes, one cover slide with cell monolayer was treated with 0.1% (v/v) DMSO in fresh medium, and another cover slide was treated with fresh medium for 24 h, when rapamycin was added to a final concentration of 50 µg/mL. After 48 h of incubation of cells with 1, DMSO (control for 1) or Rapamycin (positive control), medium was removed and cells were stained according to the instructions of CYTO-ID Autophagy detection kit (ENZO, NY, USA). Stained cells were imaged using a fluorescence microscope (Carl Zeiss) and autophagy measured using the CYTO-ID® Green dye (excitation and fluorescence emission at 463 and 534 nm, respectively; DAPI was used to counterstain the nucleus (excitation and fluorescence emission at 358 and 461 nm, respectively) and respective software (ZEN Blue edition, 2011). The total number of cells and the number of cells with autophagolysosomes were counted in at least 5 different images of each sample to calculate the % of cells in autophagocytosis.
Three-dimensional graphene oxide-coated polyurethane foams beneficial to myogenesis
Published in Journal of Biomaterials Science, Polymer Edition, 2018
Yong Cheol Shin, Seok Hee Kang, Jong Ho Lee, Bongju Kim, Suck Won Hong, Dong-Wook Han
The myogenic differentiation of C2C12 skeletal myoblasts was analyzed by immunofluorescence staining for myosin heavy chains (MHCs). The C2C12 myoblast-cultured PU and GO-PU foams were fixed with a formalin solution (3.7% formaldehyde in DPBS, Sigma-Aldrich Co.) for 10 min, and permeabilized in 0.1% of a Triton X-100 solution (Sigma-Aldrich Co.) for 5 min. The cells were then blocked in a 2% of bovine serum albumin solution (BSA, GenDEPOT, Barker, TX) for 30 min, and incubated with Alexa Fluor 488-conjugated anti-MHC monoclonal antibody (clone MF20, 1:200, eBioscienceInc., San Diego, CA) overnight at 4 °C. After immunofluorescence staining for MHCs, the cells were incubated with tetramethyl rhodamine isothiocyanate (TRITC)-conjugated phalloidin (at 1:40 in DPBS; Molecular Probes, Eugene, OR, USA) at room temperature for 20 min, followed by incubation with 4′,6-diamidino-2-phenylindole (DAPI, 1 μM, Sigma-Aldrich Co.) to counterstain the nuclei.