Liver Microcirculation
John H. Barker, Gary L. Anderson, Michael D. Menger in Clinically Applied Microcirculation Research, 2019
Acridine orange (1 µmol/kg, Sigma, St. Louis) is given as fluorescence marker of leukocytes, allowing assessment of sinusoidal perfusion and leukocyte endothelial interactions.69 For determination of red blood cell velocity, ex vivo FITC-labeled erythrocytes are used, as originally described by Zimmerhackl et al. in a modified technique.70 The labeled cells are stored at +4°C after addition of citrate-phosphate-dextrose (1.4:10) up to 5 days. Two min prior to liver microscopy, 0.05 ml red cells, 1:1 diluted with normal saline, are injected into the isogenic recipient animals. For investigation of macrophage function, fluorescence-labeled latex particles of approximately 1-µm diameter (Polysciences) are injected intravenously, as described earlier.39,67
Microbiological Diagnosis of Parasitic Diseases
Nancy Khardori in Bench to Bedside, 2018
microscopic examination of 50 to 60 pL of blood specimen after a process of micro-hematocrit centrifugation in QBC malaria tubes. These tubes are coated with a fluorescent stain (acridine orange) and certain anticoagulants. The method further involves high speed centrifugation of the blood specimen in QBC tubes. This causes density-based differential separation of different blood cells, and parasitized and normal cells, which become concentrated in different layers. The fluorescence imparted to the parasites by acridine orange dye, further helps to increase the sensitivity of the test. The sensitivity of the assay has been reported to be in a wide range of 55% to more than 90% compared to thin and thick blood film microscopy. The major drawbacks associated with this method are (1) high cost of disposables and equipment, (2) difficulty in species identification, (3) difficulty in identifying mixed infections, and (4) difficulty in determination of parasite load (Hänscheid 1999). This method may also be used to demonstrate other blood stream infections, such as Filariasis and tick-borne relapsing fever, with high level of sensitivity (Wang 1998, Van Dam et al. 1999).
Cytology of Bladder Cancer
George T. Bryan, Samuel M. Cohen in The Pathology of Bladder Cancer, 2017
Quantitative methods for measuring nucleic acids have occupied the major attention of investigators utilizing flow cytometry. Acridine orange, acriflavine Feulgen, chromomycin, and ethidium bromide are a few of the DNA or DNA and RNA fluorochromes used in cytometry. Collste and co-workers102 have made a thorough study of acridine orange and its reactions with DNA and RNA, the complexities of which are just beginning to be better understood. They have also applied their methods to bladder irrigation samples with some success. Using multiparameter determinations, they found two parameters that were needed to identify patients with bladder carcinoma: increase in proportion of urothelial cells with more than diploid DNA and aneuploid peaks.102 Tribukait et al.68,69 have used a modification of ethidium bromide staining to reduce nonspecific binding of that fluorochrome and have quantitated DNA in cells from bladder tumor tissue and from bladder washings using a rapid-flow cytofluorometer. They found increasing aneuploidy related to higher-grade tumors.
Impact of bee venom and melittin on apoptosis and biotransformation in colorectal carcinoma cell lines
Published in Toxin Reviews, 2021
Danijela D. Nikodijević, Milena G. Milutinović, Danijela M. Cvetković, Maja Đ. Ćupurdija, Milena M. Jovanović, Ivan V. Mrkić, Marija Đ. Jankulović-Gavrović, Snežana D. Marković
Phosphate-buffered saline (PBS) and Dulbecco’s modified Eagle medium (DMEM) were obtained from GIBCO (Invitrogen, Carlsbad, CA). Ethidium bromide (EB), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), absolute ethanol and chloroform (molecular biology grade) were obtained from SERVA (Heidelberg, Germany). Acridine orange (AO) was obtained from Acros Organics (Morris Plains, NJ). qPCR Kit (Luna Universal qPCR Master Mix Kit) was from BioLabs, New England (Ipswich, MA). Nuclease-free water and TRIzol were from Ambion (Austin, TX). The secondary antibody conjugated with Cy3, diamidino-2-phenylindole (DAPI) and kit for translating RNA into complementary DNA (high-capacity cDNA reverse transcription kits) from Thermo Scientific (Waltham, MA). Polyvinyl alcohol mounting medium was obtained from Fluka Analytical (Buchs, Switzerland). All solvents and chemicals were of analytical grade. Melittin (dry powder) was obtained from Sigma-Aldrich (St. Louis, MO).
Selenium and L-carnitine protects from valproic acid-Induced oxidative stress and mitochondrial damages in rat cortical neurons
Published in Drug and Chemical Toxicology, 2022
Ahmad Salimi, Nasrin Alyan, Nasim Akbari, Zhaleh Jamali, Jalal Pourahmad
The isolated neurons lysosomal membrane damage was assessed from the redistribution of the lipophilic dye acridine orange. Cells were exposed with different concentrations of VPA (50, 100 and 200 µM) or IC50 3 h VPA 1 μM SS and IC50 3 h VPA +1 mM LC for 1, 2 and 3 h in normal medium or medium at 70% confluence. SS and LC were added to medium 30 min before exposure with VPA. After the exposure time, medium was exchanges with acridine orange containing medium. The cells were incubated with acridine orange (5 µM) for 10 min and then the medium was removed. The fluorescence intensity of acridine orange was measured at the excitation wavelength of 470 nm and the emission wavelength of 540 nm by fluorescence spectrophotometer (Shimadzu RF5000U, Tokyo, Japan) (Boya and Kroemer 2008). The formula ΔF = F−F0/F0 × 100 was used for calculating of % lysosomal damage (ΔF = fluorescence intensity changes, F = fluorescence intensity and F0 = baseline fluorescence).
Bixin Triggers Apoptosis of Human Hep3B Hepatocellular Carcinoma Cells: An Insight to Molecular and IN SILICO Approach
Published in Nutrition and Cancer, 2018
Yogesh Kumar, Alugoju Phaniendra, Latha Periyasamy
AO/EB staining of treated Hep3B cell showed smilar result. Acridine orange is taken up by both viable and nonviable cells but Ethidium bromide is taken only by nonviable cells. The viable cells emits green fluorscence since acridine orange gets intercalated into double stranded nucleic acid. Nonviable or necrotic cell emits red fluorscence. Cells undergoing apoptosis fluorscence yellowish to orange due to uptake of both the dyes (Fig. 3A). Further, DNA morphological study with 4′,6-diamidino-2 phynylindole (DAPI) staining viewed under fluorescence microscope revealed that Hep3B cells treated with IC50 concentration of bixin drives cells for a phase change from a heterogenous nucleus to highly condensed, fragmented and packed into tiny bodies indicating that the cells were under apoptotic process as compared to control (Fig. 3B). Moreover, Fast halo assay also confirm the DNA damage at the single cell level (Fig. 4).
Related Knowledge Centers
- DNA
- Dye
- Fluorescein
- Fluorescence
- Nucleic Acid
- Organic Compound
- Rna
- Lysosome
- Phagocytosis
- Fluorescence Microscope