Computer Scoring of Micronuclei in Human Lymphocytes

Callisen, Hannes; Norman, Amos; Pincu, Madeleine

doi:10.1007/978-3-642-69334-2_16

Cited by 6 publications

(5 citation statements)

References 1 publication

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“…The lower limit for the detection of micronuclei by flow cytometry is therefore caused by the infrequency of very small micronuclei and the increasing frequency of debris with very low fluorescence intensities. This lower limit agrees well with microscopic observations of the size distribution of micronuclei in mouse bone marrow cells (11) or in human lymphocytes (3,24). By image analysis of the size distribution of micronuclei, the smallest micronuclei that can be detected also have relative DNA contents of about 0.5-1% of the main nucleus (8).…”

Section: Discussionsupporting

confidence: 87%

“…Even DNA synthesis in radiationinduced micronuclei has been demonstrated using incorporation of bromodeoxyuridine (BrdUrd) and anti-BrdUrd antibodies (15,181. Usually, microscopic scoring of micronuclei in some hundreds of cells is a time-consuming procedure. Therefore several attempts have been made to automate micronucleus scoring by image analysis (3,8,24,25,27) or by flow cytometric techniques (9,17,21,28). Flow cytometry seems to be a n especially powerful technique to analyse rapidly large numbers of cells.…”

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confidence: 99%

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Multiparametric flow cytometric analysis of radiation‐induced micronuclei in mammalian cell cultures

Schreiber

Beisker²,

Bauchinger³

et al. 1992

Cytometry

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A new flow cytometric method is presented that quantifies the frequency of radiation-induced micronuclei in mammalian cell cultures with high precision. After preparing a suspension of main nuclei and micronuclei stained with ethidium bromide and Hoechst 33258, both types of particles are measured simultaneously in a flow cytometer using forward light scatter and three fluorescence emission intensities excited by UV, 488 nm, and by energy transfer from Hoechst 33258 to ethidium bromide. Nonspecific debris overlapping the micronucleus distribution especially in the low fluorescence intensity region was discriminated from micronuclei by calculating ratios of the different fluorescences. The frequencies of radiation-induced micronuclei measured with this new technique agreed well with results obtained by conventional microscopy. The lower limit of the DNA content of micronuclei identified by this technique was found to be about 0.5YA.75% of the DNA content of GI-phase nuclei. Dose effect curves and the time-dependent induction of micronuclei were measured for two different mouse cell lines.

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Section: Discussionsupporting

confidence: 87%

mentioning

confidence: 99%

Multiparametric flow cytometric analysis of radiation‐induced micronuclei in mammalian cell cultures

Schreiber

Beisker²,

Bauchinger³

et al. 1992

Cytometry

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“…Thus, it is possible to automate the image processing, avoiding the tiresome work of scoring. Image processing is performed by specially designed computers for image analysis (1) or by image processing systems based on personal computers (PCs) with appropriate hardware accessories [e.g., video digitizer cards and image processing boards (2)(3)(4)]. Our present work deals with the design and development of such a system.…”

Section: Introductionmentioning

confidence: 99%

Computerized image analysis for determining micronucleus frequency.

Szirmai¹,

Bérces²,

Köteles³

1993

Environ Health Perspect

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A method for the computerized automation of micronucleus scoring is presented. The task is to identify the cultured, cytokinesis-blocked peripheral lymphocytes (CB cells) and their micronuclei (MN). The main parts of the hardware are the video camera attached to the microscope, the IBM-compatible personal computer with the video digitizer card, and the computer-controlled stage movement unit. The computerized image processing is based on determination and interpretation of contour lines of the CB cells, nuclei, and MN. The BNCTEST image processing software has been developed up to the demonstration phase, and now it has been prepared for the testing period of image series on a large scale.ImagesFIGURE 1.

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“…There is a growing list of pollutants known to induce a high incidence of micronuclei in laboratory conditions (Hooftman and Raat 1982;Das and Nanda 1986;Hughes and Hebert 1991). Callisen et al (1984) were the first to demonstrate quantitation of nuclei and micronuclei using CIA. The modified differential staining technique of Fenech and Morley (1985) used in the present work made the recognition of micronuclei and nuclei easier and the observed volume relation could be attributed to formation of micronuclei.…”

Section: Resultsmentioning

confidence: 99%

“…Since early attempts to automate the scoring of micronuclei using flow cytometry (Hutter and Stoehr 1982;Leary andTometsko 1986 cited in Romagna andStaniforth 1989) and semi-automated image analysis (Andrese et al 1986 cited in Romagna and Staniforth 1989), several investigators have attempted to develop computer software for micronucleus detection. While, Callisen et al (1984) demonstrated the quantitation of nuclei and micronuclei by computerized image analysis (CIA), Romagna and Staniforth (1989) developed a new method of removing nucleated bone marrow cells in rodents, using a cellulose column. Although acceptable, the methodology is labour intensive and not suited for large volume testing.…”

Section: Introductionmentioning

confidence: 99%

Computerized automated morphometric assay including frequency estimation of pentachlorophenol induced nuclear anomalies (micronucleus) in catfish Heteropneustes fossilis

Ahmad¹,

Ali²,

Farah³

et al. 2002

Chromosoma

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An in vivo study of the effects of pentachlorophenol was carried out with a pre-acclimatized fish species, Heteropneustes fossilis, using four sub-lethal concentrations, 0.1, 0.2, 0.3 and 0.4 ppm, and three sampling times, 48, 72 and 96 h. Cytogenetic preparations were stained by the haematoxylin-eosin technique. The incidence of micronuclei was scored by a manual and an automated method. Small-sized micronuclei appeared in the cytoplasm in addition to the main nucleus. The frequency of micronucleated erythrocytes peaked at 4 days (96 h) exposure. The percentage of single micronuclei increased with longer exposures. The Mann-Whitney U test showed all micronuclei frequencies were significantly different from control (P<0.05). No statistical difference was observed between scores obtained by the manual and automated methods. A linear relationship between the percentage of micronucleated erythrocytes and dose was confirmed at all levels. Computer image analysis of morphological variations of erythrocytes indicated a 1:5 ratio of micronuclei and main nucleus accompanied by a reduction in cell volume by 600 dot units. Pentachlorophenol-mediated genotoxicity was confirmed in this fish for the first time. Possible consequences of genotoxicity and cytotoxicity are discussed.

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Computer Scoring of Micronuclei in Human Lymphocytes

Cited by 6 publications

References 1 publication

Multiparametric flow cytometric analysis of radiation‐induced micronuclei in mammalian cell cultures

Multiparametric flow cytometric analysis of radiation‐induced micronuclei in mammalian cell cultures

Computerized image analysis for determining micronucleus frequency.

Computerized automated morphometric assay including frequency estimation of pentachlorophenol induced nuclear anomalies (micronucleus) in catfish Heteropneustes fossilis

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