A fluorescent complex between ethidium bromide and nucleic acids

LePecq, Jean-Bernard; Paoletti, Claude

doi:10.1016/0022-2836(67)90353-1

Cited by 2,324 publications

(918 citation statements)

References 26 publications

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“…It is well known that Na + can partly neutralize the phosphate group of DNA; if the interaction between LaF 3 colloids and DNA is electrostatic, the quenched fluorescent intensity will be weakened by adding NaCl (26). The results showed that the addition of NaCl had no significant effect on the fluorescence intensity of the system.…”

Section: The Interaction Of Laf 3 -Poly(st-co-maa) Colloids With Dnamentioning

confidence: 94%

Luminescent and hydrophilic LaF₃–polymer nanocomposite for DNA detection

Wang

2008

Luminescence

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Luminescent LaF 3 -Ce 3+ /Tb 3+ nanocrystals have been successfully prepared via a simple wet chemical technique. For the next bioapplication, these nanoparticles dispersed in cyclohexane have also been functionalized with poly(Stco-MAA), based on a designed oil-in-water microemulsion system. These polymer-coated nanospheres are water-soluble and bioconjugable. Unlike semiconductor quantum dots, the as-prepared lanthanum fluoride nanocrystals possess nonsize-dependent emissions and completely stable photocycles. With functionalized LaF 3 nanospheres as fluorescence probes, a fluorescence method was developed for the rapid quantitative analysis of DNA, due to the quenching effect of fluorescence by the DNA. Under optimum conditions, the fluorescence intensity was proportional to the concentration of the introduced DNA over the range 2.5-35 mg/mL for calf thymus DNA (ctDNA) and 2.5-30 mg/mL for fish sperm DNA (fsDNA), respectively.

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Section: The Interaction Of Laf 3 -Poly(st-co-maa) Colloids With Dnamentioning

confidence: 94%

Luminescent and hydrophilic LaF₃–polymer nanocomposite for DNA detection

Wang

2008

Luminescence

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“…This was caused by a dilution of staining solution with sheath liquid in the tubing. Because of a higher ionic strength of the saline the binding of the PI was reduced (7,8) until the tubing was cleared completely of saline. The problem was solved by using stock solution as sheath liquid.…”

Section: Methodsmentioning

confidence: 99%

A Detergent‐trypsin method for the preparation of nuclei for flow cytometric DNA analysis

Vindeløv¹,

Christensen²,

Nissen³

1983

Cytometry

1,526

655

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A new modification of our detergent technique for the preparation of nuclei for flow cytometric DNA analysis is described. The attainment of low coefficients of variation of the peaks and of quantitative staining of nuclei from different tissues was a problem with the original method. This was solved in the new modification by trypsinization of the unfixed nuclei. The nuclei were stabilized by spermine. A simple procedure for long-term storage of samples at -80°C was integrated into the method. The fluorescence of the nuclei was stable for at least 3 hours after staining. Light exposure protection of the samples was essential. No cell loss was caused by storage or staining. The method was successfully applied on samples including: (a) Normal tissues-human lymphocytes, granulocytes and spleen. Mouse lymphocytes, bone marrow, spleen, liver, kidney and thymus. (b) Human neoplasms-lung cancer, breast cancer, lymphoma, leukemia, bladder cancer and cancer of the oral cavity. Key terms: Flow cytometry, DNA, propidium iodide, solid tissues, preparation, storage A previously published method for the preparation of fineneedle aspirates from solid tissues for flow cytometric DNA analysis (9) has been useful for examination of human solid tumors (1,13,14). The method is based on detergent lysis of cell membranes and produces stained nuclei in monodisperse suspension in a single step. However, the low salt procedure of the technique, which has been used most extensively, has two major drawbacks: (a) Tissues rich in cytoplasm, for instance liver cells, are not prepared optimally because residual cytoplasm contaminates the nuclei, causing clumping and nonspecific staining. In the DNA distribution this produces asymmetrical peaks with high coefficients of variation (cv). ( b ) Staining of different types of cells is not perfectly quantitative. Thus a comparison of human lymphocytes and granulocytes for example showed a 6% difference in fluorescence.The original method was therefore modified with the aim of obtaining a preparation technique that would produce optimal results in a broad spectrum of tissues in regard to

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“…This procedure involved the suspension of the cells in a detergent based staining solution such that the nuclei were stripped of cytoplasm. Since PI stains double stranded nucleic acid (16) and ribonuclease was present, the resulting fluorescence was proportional to DNA. Results PY staining specificity in EL 4 tumor cells: Figure 1 illustrates the staining specificity of PY to EL4 cells.…”

Section: Methodsmentioning

confidence: 99%

Flow cytometric analysis of RNA content in different cell populations using pyronin Y and methyl green

et al. 1982

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Pyronin Y (PY) was used, in flow cytometric (FCM) systems, to estimate the RNA content per cell in formalin fixed EL4 leukosis tumor cells, enzyme dispersed R3327-G rat prostatic adenocarcinoma cells, mouse spleen cells stimulated with concanavalin A, and human peripheral blood lymphocytes stimulated with phytohemagglutinin. Preincubation of the cells with methyl green (MG) blocked PY binding to DNA such that the intracellular fluorescence from MG-PY was due primarily to its binding to RNA. Treatment of the cells with ribonuclease resulted in a 3-to &fold reduction in the fluorescence intensity of intracellular MG-PY.Mitogen stimulation of either mouse or human lymphocytes resulted in an increase in DNA (propidium iodide fluorescence) and RNA (MG-PY fluorescence) content per cell over resting levels. Further, the changes in stimulated human lymphocyte DNA and RNA contents following 24,48, and 72 hr of cell culture were monitored, The results showed that RNA levels were significantly increased prior to that of DNA. Also, the effects of different cell cycle phase specific blocking agents on lymphocyte cell cycle traverse were investigated. We found that: a) actinomycin D inhibited the increases in cellular RNA and DNA; b) hydroxyurea inhibited the increases in cellular DNA whereas increases in cellular RNA were only slightly reduced; c) tritiated thymidine caused an accumulation of cells having high DNA and RNA contents; and d) Colcemid promoted an accumulation of cells having high DNA contents while causing a reduction of cells having high RNA contents. These results were nearly identical to reports by other investigators using the metachromatic dye acridine orange to quantitate RNA per cell. Thus, the MG-PY technique described is indicated to provide a stable and accurate measure of RNA content per cell.Key terms: Pyronin Y, methyl green, flow cytometry, RNA Over the last decade the automated identification and classification of cellular subpopulations has developed extensively using flow cytometric (FCM) instrumentation. This instrumentation has been particularly useful in the rapid discrimi-'

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A fluorescent complex between ethidium bromide and nucleic acids

Cited by 2,324 publications

References 26 publications

Luminescent and hydrophilic LaF₃–polymer nanocomposite for DNA detection

Luminescent and hydrophilic LaF₃–polymer nanocomposite for DNA detection

A Detergent‐trypsin method for the preparation of nuclei for flow cytometric DNA analysis

Flow cytometric analysis of RNA content in different cell populations using pyronin Y and methyl green

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A fluorescent complex between ethidium bromide and nucleic acids

Cited by 2,324 publications

References 26 publications

Luminescent and hydrophilic LaF3–polymer nanocomposite for DNA detection

Luminescent and hydrophilic LaF3–polymer nanocomposite for DNA detection

A Detergent‐trypsin method for the preparation of nuclei for flow cytometric DNA analysis

Flow cytometric analysis of RNA content in different cell populations using pyronin Y and methyl green

Contact Info

Product

Resources

About

Luminescent and hydrophilic LaF₃–polymer nanocomposite for DNA detection

Luminescent and hydrophilic LaF₃–polymer nanocomposite for DNA detection