Analysis of Plasmodium falciparum growth in culture using acridine orange and flow cytometry.

Hare, J.D.; Bahler, David W.

doi:10.1177/34.2.2418101

Cited by 37 publications

(26 citation statements)

References 19 publications

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“…However, they require flow cytometers equipped with ultraviolet lasers that are not available in most laboratories. Nonspecific DNA staining agents, such as thiazole orange (7), acridine orange (8)(9)(10), ethidium bromide (11), propidium iodide (12), or hydroethidine (13)(14)(15), have also been used to stain nucleic acids from infected erythrocytes. Recently, YOYO-1, a cyanine dimer, has been added to this list (16).…”

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confidence: 99%

Improvement of detection specificity of Plasmodium‐infected murine erythrocytes by flow cytometry using autofluorescence and YOYO‐1

et al. 2005

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Background: Microscopic analysis of blood smears is currently the most frequently used method to measure parasitemias in experiments of drug efficacy in murine models of malaria. However, it is subjective and labour intensive, which preclude its utilization in large-scale evaluation programs. Flow cytometry is an alternative method, but due to the limited specificity achieved with the currently available techniques, it has not been widely used in murine models of malaria during preclinical evaluation. We describe a new flow cytometric method based on the differences of autofluorescence and DNA content measured after staining with YOYO-1 that are observed in infected erythrocytes compared with noninfected erythrocytes. Methods: Samples of blood from Plasmodium yoeliiinfected animals were fixed with glutaraldehyde, incubated with RNAase, and stained with YOYO-1 in 96-well plate format. After acquisition, erythrocytes gated in loga-

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confidence: 99%

Improvement of detection specificity of Plasmodium‐infected murine erythrocytes by flow cytometry using autofluorescence and YOYO‐1

et al. 2005

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“…The labelling of parasitized RBCs (PRBC) was performed in the dark without permeabilization of the cells in two steps [10,13,31], using two nucleic acids staining: i) vital dye hydroethidine (HE) (Interchim 17084), which is metabolized into ethidium by esterases in intact PRBC (ethidium labelling of nucleic acids results in a red fluorescence emission) [11] (Figure 1B); and, ii) thiazole orange (TO) (Sigma 17237), which binding both to RNA and DNA emitting a green fluorescence [15]. HE is prepared at 10 mg/ml in dimethyl sulphoxide then stored at −20°C.…”

Section: Methodsmentioning

confidence: 99%

“…To counteract these difficulties, several dyes have been used to measure division of the nucleus by fluorimetry [8] or by flow cytometry. The most often used are: Hoechst 33258 [9], acridine orange [8,10,11], thiazole orange [12], hydroethidine [13], and recently YOYO-1 [5,14-16]. Sybergreen I based test was also standardized and is currently used in several laboratories [7].…”

Section: Introductionmentioning

confidence: 99%

Cytometric measurement of in vitro inhibition of Plasmodium falciparum field isolates by drugs: a new approach for re-invasion inhibition study

Varela

Razakandrainibe

Aldebert³

et al. 2014

Malar J

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BackgroundA flow cytometric method is proposed to study in vitro drug sensitivity of Plasmodium falciparum. Standard [3H]-hypoxanthine incorporation assay gives only information on inhibition of maturation by drugs. This method is usable on field isolates and provides data on both inhibition of maturation and re-invasion.MethodsThe method is based on the staining of parasites with hydroethidine (HE) and thiazole orange (TO) which allow differential identification of early, trophozoite and late stage of the parasite by flow cytometry. Late stages of the parasites are obtained by incubation in culture for 24 hours. Reinvasion is followed by culturing parasitized red blood cells for 24 h more.ResultsCompared to the standard [3H]-hypoxanthine incorporation assay, it gave similar results as expressed by 50% inhibitory concentrations for chloroquine of laboratory strains and “field” isolates. The effect of quinine on the schizont-ring transition was also explored using this method. First data on the inhibition of re-invasion induced by quinine are presented for both P. falciparum-cultured strains and field isolates.DiscussionThis method is simple to use event for field isolate study. It is suitable to analyse effect of drugs on steps of the parasite life cycle different for the maturation one. Using this method quinine was found to have a inhibitory effect on re-invasion of red cells by Plasmodium.

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“…Assess ment of RNA content in archival material is more difficult, however, and previous stud ies have noted a number of artifacts [18]. We and others [19][20][21] have shown that acridine orange selectively stains RNA in formalinfixed cells. Intrasample and daily variability in nuclear RNA measurements were within experimental error.…”

Section: Dnamentioning

confidence: 99%

Flow Cytometry Measurement of Nuclear RNA Content in Uveal Melanoma

Chen

Char²,

Waldman³

et al. 1990

Ophthalmic Res

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We retrospectively studied the nuclear RNA content in uveal melanomas with known, long-term follow-up. Thirty patients had spindle cell melanomas, 25 had mixed cell tumors, and 9 had epithelioid cell neoplasms. Epithelioid cell types had a higher nuclear RNA content compared to mixed or spindle cell types (p < 0.05). Using the Cox proportional hazards model, the nuclear RNA content appeared to be an independent prognostic indicator in uveal melanomas, and an increased nuclear RNA content was associated with a worse prognosis (p = 0.001).

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Analysis of Plasmodium falciparum growth in culture using acridine orange and flow cytometry.

Cited by 37 publications

References 19 publications

Improvement of detection specificity of Plasmodium‐infected murine erythrocytes by flow cytometry using autofluorescence and YOYO‐1

Improvement of detection specificity of Plasmodium‐infected murine erythrocytes by flow cytometry using autofluorescence and YOYO‐1

Cytometric measurement of in vitro inhibition of Plasmodium falciparum field isolates by drugs: a new approach for re-invasion inhibition study

Flow Cytometry Measurement of Nuclear RNA Content in Uveal Melanoma

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