Does acridine orange stain dead cells?

Does acridine orange stain dead cells?

Acridine orange is an intercalating dye that can permeate both live and dead cells. Propidium iodide can only enter dead cells with poor membrane intergrity, so it will stain all dead nucleated cells to generate red fluorescence.

How does acridine orange work?

Acridine orange is an organic compound that serves as a nucleic acid-selective fluorescent dye with cationic properties useful for cell cycle determination. Acridine orange is cell-permeable, which allows the dye to interact with DNA by intercalation, or RNA via electrostatic attractions.

How is cell apoptosis measured?

Apoptosis is detected by measuring the externalization of phosphatidylserine on the plasma membrane using fluorescent-tagged annexin V. Additionally, flow cytometry can be employed to determine alterations in cell size (Bortner and Cidlowski, 2001; Warnes et al. 2011).

Can ethidium bromide stain live cells?

Ethidium bromide will stain only cells that have lost membrane integrity. Live cells will appear uniformly green. Early apoptotic cells will stain green and contain bright green dots in the nuclei as a consequence of chromatin condensation and nuclear fragmentation.

What is the main problem with acridine orange stain?

Since it is a cationic dye, it also enters acidic compartments such as lysosomes which in low pH conditions, will emit orange light. Acridine orange is a carcinogen when absorbed through the skin. Wear gloves when working with this stain.

What is acridine orange staining?

Acridine Orange (AO) is a nucleic acid selective metachromatic stain useful for cell cycle determination. AO interacts with DNA and RNA by intercalation or electrostatic attraction respectively. This procedure optimally stains cells for analysis by flow cytometry.

Is acridine orange soluble in water?

This product is soluble in water (6 mg/ml), in ethanol (2 mg/ml), and in 2-methoxyethanol (EGME, 4 mg/ml). 1 At 1 mg/mL in water, it forms a clear, dark orange to amber solution. 1. The Sigma-Aldrich Handbook of Stains, Dyes & Indicators, Green, F.J., Ed., Aldrich Chemical Co.

How does acridine orange bind?

Acridine Orange is a cell-permeant nucleic acid binding dye that emits green fluorescence when bound to dsDNA and red fluorescence when bound to ssDNA or RNA. This unique characteristic makes acridine orange useful for cell-cycle studies. Acridine orange has also been used as a lysosomal dye.

What are the markers of apoptosis?

Potentially useful apoptosis markers include cleaved cytokeratin-18 (c-CK18), cleaved caspase-3 (c-cas-3), cleaved lamin A (c-lam-A), phosphorylated histone H2AX (gammaH2AX), cleaved poly(ADP ribose) polymerase (c-PARP), and translocation of apoptosis-inducing factor (AIF).

How do you assay for apoptosis?

There are a number of methods for running an apoptosis assay to measure these markers of apoptosis.

1. Annexin V binding of cell surface phosphotidylserine.
2. DNA condensation and fragmentation (TUNEL) assays.
3. Caspase activation and detection assays.
4. Mitochondrial membrane potential-dependent dyes.
5. Cytochrome C release assays.

How do you make acridine orange solution?

Preparation of reagent: 50 mg acridine orange is dissolved in 10 ml of distilled water to prepare a stock solution and stored in the refrigerator. 1 ml of acridine orange stock solution and 0.5 ml of glacial acetic acid is added to 50 ml of distilled water to prepare a working solution.

How does EtBr cause mutations?

Loss of intercalating agent can result in deletion. Intercalating agents introduce atypical spacing between base pairs, resulting in DNA polymerase introducing either a deletion or an insertion, leading to a potential frameshift mutation.

Can acridine orange/ethidium bromide staining detect apoptosis in tumor cells?

Our results suggest that dual acridine orange/ethidium bromide staining is an economic and convenient method to detect apoptosis in tumor cells and to test tumor chemosensitivity compared with flow cytometry.

What is acridine orange/ethidium bromide staining?

Acridine orange/ethidium bromide (AO/EB) staining is used to visualize nuclear changes and apoptotic body formation that are characteristic of apoptosis.

Can AO/EB staining detect apoptosis in HepG2 cells?

In this study, BS-induced apoptotic cell death in HepG2 cells. AO/EB staining is a gold standard technique to detect cells undergoing apoptosis (Kasibhatla et al., 2006; Liu et al., 2015;Shebi et al., 2019).

Can AO/EB staining detect apoptosis in osteosarcoma cells?

Dual AO/EB staining to detect apoptosis in osteosarcoma cells compared with flow cytometry Our results suggest that dual acridine orange/ethidium bromide staining is an economic and convenient method to detect apoptosis in tumor cells and to test tumor chemosensitivity compared with flow cytometry.