Flow cytometric S‐phase fraction measurement in breast carcinoma: Influence of software and histogram resolution

Jourdan, Marie-Lise; Ferrero-Poüs, Magali; Spyratos, Frédérique; Romain, Sylvie; Martin, Pièrre‐Marie; Chassevent, Agnès

doi:10.1002/cyto.10116

Cited by 18 publications

(12 citation statements)

References 29 publications

(41 reference statements)

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“…The evaluation of DNA ploidy, by measuring the FCM DI, has shown its prognostic value in childhood ALL (Look et al, 1985;Pui et al, 1991;Maloney et al, 2000;Raimondi et al, 2006;Schultz et al, 2007;Aricò et al, 2008). Its use in therapeutic strategy has been justified by work carried out by American (Look, Roberson & Murphy, 1987;Pui et al, 1991;Trueworthy et al, 1992) and European (Kaspers et al, 1995;Aricò et al, 2008) standardized approach made results comparable from several teams with different FCM systems, including solid tumors studies, where the difficulty is increased in comparison with hematological pathologies (D'Hautcourt, Spyratos & Chassevent, 1996;Chassevent et al, 2001;Jourdan et al, 2002). In any case, different technical precautions are essential to obtain interpretable results as, for example, the need for preparatory methods that allow the optimal quantification of DNA content and of course the correct elimination of aggregates.…”

Section: Discussionmentioning

confidence: 99%

DNA Index in childhood acute lymphoblastic leukaemia: a karyotypic method to validate the flow cytometric measurement

Rachieru-Sourisseau

Baranger

Dastugue

et al. 2010

Int J Lab Hematology

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The DNA index (DI) is a prognostic factor in childhood acute lymphoblastic leukemia (ALL). The accuracy of DI measurement is important for treatment stratification: hyperdiploidy with DI > or = 1.16 is predictive of favorable prognosis whereas hypodiploidy is associated with poor prognosis. The aim of this study was to validate the accuracy of the DI measured by flow cytometry (FCM) by comparison with the karyotype. From samples of 112 childhood ALL, we created a formula to calculate a theoretical DNA index (tDI) based on the blast cell karyotype, taking into account the additional or missing chromosome material of the major clone. FCM DI correlated with tDI calculated from karyotype (R = 0.987) and with modal chromosome number (DI = 0.0202 x Modal NB + 0.0675 and R = 0.984). In three cases a hypodiploid blast cell population was detected by FCM, while only the duplicated clone was identified by the karyotype. The strong correlation between tDI and DI validates the accuracy of FCM quantification, which is technically fast on fresh or frozen samples. If the karyotype is essential to analyze chromosomal abnormalities, FCM provides complementary information in aneuploid ALLs, either by confirming the cytogenetic data or by detecting additional clones not identified when only using cytogenetic data.

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Section: Discussionmentioning

confidence: 99%

DNA Index in childhood acute lymphoblastic leukaemia: a karyotypic method to validate the flow cytometric measurement

Rachieru-Sourisseau

Baranger

Dastugue

et al. 2010

Int J Lab Hematology

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“…6. Cell cycle analysis by mathematic modelling (Jourdan et al, 2002) by fitting software (e.g,. Modfit™) usually underestimates percentage of cells in S-phase, since G1 and G2/M peaks are fitted by a gaussian model and early and late S-phase are included inside fitted peaks.…”

Section: Mps-1 Kinase Inhibitor (Nms-p715)mentioning

confidence: 99%

Assessment of Cell Cycle Inhibitors by Flow Cytometry

Cappella¹,

Moll²

2011

Drug Discovery and Development - Present and Future

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“…Properly applied, today's available software allows the accurate calculation of the DNA index (DI) of the cell population of interest as well as an exact SPF calculation for up to three different populations in a given sample. Detailed comparison of different software calculation modes has revealed the impact of data handling on intra-and interlaboratory reproducibility [9,10,29,45]. These studies underline the importance of a standardized software-supported data calculation.…”

Section: Data Collection Processing and Assessmentmentioning

confidence: 91%

Flow Cytometric DNA Analysis in Oncology: From Single to Multiparametric Measurements/Durchflusszytometrische DNA-Analytik in der Onkologie: Von einzel- zu multiparametrischen Messungen

Brockhoff¹,

Knuechel²

2003

LaboratoriumsMedizin

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DNA analysis was one of the earliest applications in flow cytometry. The stoichiometric interaction of fluorescent dyes with nucleic acid allows the evaluation of DNA content (ploidy) and the proliferative fraction. Initially, DNA was measured as the only parameter along with side-and forward-scatter properties to identify intact nuclei or cells. Later, a dual-parametric approach for the simultaneous analysis of DNA and protein was introduced. Today, the main application of flow cytometric DNA analysis is the additional, tumour entity-dependent diagnosis in pathology. The knowledge about an abnormal DNA content as a crude indicator of chromosomal instability and high S-phase fraction can result in a precise determination of tumour prognosis and can lead to the optimization of individual therapy and complementary disease monitoring. DNA analysis is applied in both haematological and solid tumour diagnosis. In contrast to haematological diseases, the analysis of solid tumours requires the isolation of cell nuclei or cell disaggregation with preservation of cell integrity. The uniparametric investigation of cell nuclei in suspension allows high-resolution DNA analysis with detection of aneuploidy in the neardiploid range. However, the multiparametric approach on the basis of cell selection or discrimination takes tumour heterogeneity into account, and DNA assessment can be combined with antigen detection with additional cell phenotyping. In contrast to unimodal DNA analysis, the multiparametric approach allows a more accurate calculation of cell-cycle fractions of individual cell subpopulations. All approaches require adequate standardization regarding cell preparation, data acquisition and handling to ensure lab-independent DNA data interpretation and utilization.

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Flow cytometric S‐phase fraction measurement in breast carcinoma: Influence of software and histogram resolution

Cited by 18 publications

References 29 publications

DNA Index in childhood acute lymphoblastic leukaemia: a karyotypic method to validate the flow cytometric measurement

DNA Index in childhood acute lymphoblastic leukaemia: a karyotypic method to validate the flow cytometric measurement

Assessment of Cell Cycle Inhibitors by Flow Cytometry

Flow Cytometric DNA Analysis in Oncology: From Single to Multiparametric Measurements/Durchflusszytometrische DNA-Analytik in der Onkologie: Von einzel- zu multiparametrischen Messungen

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