Intercellular Communication in Three-Dimensional Culture
Rolf Bjerkvig in Spheroid Culture in Cancer Research, 2017
Spheroids, heart fragments, and confrontation cultures were washed in PBS and fixed in Bouin solution for 2 h at room temperature. Cell cultures were then dehydrated in 70% ethanol for 2 to 3 h, 80 and 96% isopropanol for 30 min at room temperature, and in 100% isopropanol (replaced for three times) for 90 min at 45°C. After impregnation overnight at 57°C in a mixture of equal parts isopropanol and the embedding medium Paraplast Plus (Polyscience, Warrington, PA), cell cultures were infiltrated for 2 × 7 h at 57°C with pure Paraplast Plus and finally embedded in Paraplast Plus. For routine histology all specimens were cut to 5-µm-thick sections and stained with hematoxylin/erythrosine (Chroma, Köngen, FRG).
Sexually Transmitted Diseases
Victor A. Bernstam in Pocket Guide to GENE LEVEL DIAGNOSTICS in Clinical Practice, 2019
Fixation in Bouin solution markedly reduces the rate of detection of HPV DNA by ISH compared to that in tissue fixed with buffered formalin. Fixation and processing of tissues may impede the penetration of probes to the viral DNA.
Atypical enteropathogenic E. coli are associated with disease activity in ulcerative colitis
Published in Gut Microbes, 2022
Maximilian Baumgartner, Rebecca Zirnbauer, Sabine Schlager, Daniel Mertens, Nikolaus Gasche, Barbara Sladek, Craig Herbold, Olga Bochkareva, Vera Emelianenko, Harald Vogelsang, Michaela Lang, Anton Klotz, Birgit Moik, Athanasios Makristathis, David Berry, Stefanie Dabsch, Vineeta Khare, Christoph Gasche
Fifty-seven E. coli isolates were grown on MacConkey agar for 24 hours under aerobic or anaerobic conditions, using Anaerobox and AnaeroGen sachets (Thermo Scientific, Oxoid). Single colonies were inoculated in 5 ml brain heart infusion (BHI, 37 g/L) medium with supplements (5 g/L yeast extract, 1 g/L NaHCO3, 1 g/L L-cysteine, 1 mg/L vitamin K1, 5 mg/L hemin) or in LB medium and grown under aerobic or anaerobic conditions for 6 hours at 37°C. Bacterial cells were diluted to an OD600 = 0.05. 100 µl of cell suspension was transferred to the U-bottom polystyrene 96-well plates (Costar) in four technical replicates. Plates were incubated at 37°C for 48 hours under aerobic or anaerobic conditions. Supernatants were removed, bacterial biofilms were fixed with 150 μL BOUIN solution (0.9% picric acid, 9% formaldehyde and 5% acetic acid) for 15 min and washed three times with 190 μL PBS. For staining, 150 μL 0.1% crystal violet solution was added for 10 min and washed three times with H20. For biofilm quantification, crystal violet in dried plates was dissolved in 190 μL 30% acetic acid and the plate was placed on a shaker for 1 h. Absorbance of 1:5 dilutions was measured on an Anthos 2010 microplate reader at 595 nm and 405 nm reference wavelength. Pairwise comparison was performed with the Mann–Whitney U test and ANOVA with Dunn’s multiple comparison test for comparing multiple groups.
Histological and biochemical investigation of the renoprotective effects of metformin in diabetic and prostate cancer model
Published in Toxicology Mechanisms and Methods, 2021
Pınar Koroglu-Aydın, Bertan Boran Bayrak, Ilknur Bugan, Omur Karabulut-Bulan, Refiye Yanardag
For the purpose of light microscope examinations, the kidney tissues were fixed for 22 hours in Bouin solution for routine histological procedure, processed routinely and embedded in paraffin. A 4 μm-thick sections taken at the microtome were stained using hematoxylin and eosin (HE). Semi-quantitative histological evaluation was made using an alteration of criteria defined by Fernandez et al. (2001). Each histological variable received a score between 0 (normal) and 3 (maximum damage). The scoring was done as none (−) 0, mild (+) 1, moderate (++) 2 and severe (+++) 3. These criteria were: observation of cell debris in the lumen of proximal and distal tubules, decrease in glomerular volume, presence of pycnotic nuclei, hyperemia, necrotic areas, hypertrophy and enlargement of the capsular area. Preparation samples were photographed with an Olympus CX41 microscope equipped with an Olympus DP71 digital camera DP7, Tokyo, Japan. A minimum of five area for each slide were analyzed and scored semi quantitatively for the seriousness of the changes by a blind observer.
Short-term inhalation study of graphene oxide nanoplates
Published in Nanotoxicology, 2018
Young Hun Kim, Mi Seong Jo, Jin Kwon Kim, Jae Hoon Shin, Jin Ee Baek, Hye Seon Park, Hyo Jin An, Jong Seong Lee, Boo Wook Kim, Hoi Pin Kim, Kang Ho Ahn, KiSoo Jeon, Seung Min Oh, Ji Hyun Lee, Tomomi Workman, Elaine M. Faustman, Il Je Yu
After collecting blood samples, the rats were euthanized using the anesthesiant Entobar®, followed by careful removal of the testes, heart, thymus, trachea, lungs, kidneys, spleen, liver, and brain. The organs were examined for any gross lesions, and then weighed and fixed in a 10% formalin solution containing neutral phosphate-buffered saline (PBS). For the histopathological evaluation, the testes were fixed in a Bouin solution, at killing, while the left lungs were fixed in a 10% formalin solution (BBC Biochemical, Washington, DC) containing neutral phosphate-buffered saline under 25 cm of water pressure. After fixing the organs in 10% natural PBS for 1 week, they were then embedded in paraffin and stained with hematoxylin and eosin (BBC biochemical, Washington, DC). All the animal organs were examined using light microscopy. The left lungs were sectioned into three and examined.
Related Knowledge Centers
- Acetic Acid
- Fixation
- Gastrointestinal Tract
- Histology
- Picric Acid
- Electron Microscope
- Cell Nucleus
- Formaldehyde
- Staining
- Ultrastructure