Isolation, Purification and Sorting by Flow Cytometry of Metaphase Chromosomes of Haplopappus Gracilis

A new method for decomposing flow cytometry histograms of isolated human metaphase chromosomes is described and tested. The method is based on fitting a template, composed of the means of all chromosomes of a normal karyotype to the flow histogram. The utility of the method is demonstrated by application to flow measurements of chromosomes from a normal person and comparing the results with those obtained by conventional cytophotometry. The power of the method for detecting gross chromosomal abnormalities, such as trisomy 21, as well as more subtle variations such as a single translocation, is determined for simulated data.

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Immunofluorescent detection of histone 2B on metaphase chromosomes using flow cytometry

Chromosoma ◽

10.1007/bf00287038 ◽

1984 ◽

Vol 90 (4) ◽

pp. 295-302 ◽

Cited By ~ 24

Author(s):

Barb Trask ◽

Ger van den Engh ◽

Joe Gray ◽

Marty Vanderlaan ◽

Bryan Turner

Keyword(s):

Flow Cytometry ◽

Metaphase Chromosomes

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Application of Flow Cytometry for the Isolation of Metaphase Chromosomes from Protoplasts of Nicotiana Plumbaginifolia

Progress in Plant Protoplast Research - Current Plant Science and Biotechnology in Agriculture ◽

10.1007/978-94-009-2788-9_108 ◽

1988 ◽

pp. 303-304 ◽

Cited By ~ 2

Author(s):

Harrie C. P. M. van der Valk ◽

Harrie A. Verhoeven

Keyword(s):

Flow Cytometry ◽

Nicotiana Plumbaginifolia ◽

Metaphase Chromosomes

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Strategies for choosing a deoxyribonucleic acid stain for flow cytometry of metaphase chromosomes.

Journal of Histochemistry & Cytochemistry ◽

10.1177/25.8.70463 ◽

1977 ◽

Vol 25 (8) ◽

pp. 954-964 ◽

Cited By ~ 25

Author(s):

R H Jensen ◽

R G Langlois ◽

B H Mayall

Keyword(s):

Flow Cytometry ◽

Energy Transfer ◽

Ethidium Bromide ◽

Deoxyribonucleic Acid ◽

Fluorescent Dyes ◽

Chinese Hamster ◽

Fluorescence Properties ◽

Chromomycin A3 ◽

Metaphase Chromosomes ◽

Mitotic Cells

Requirements for flow cytometry of metaphase chromosomes stained with three deoxyribonucleic acid (DNA)-specific fluorescent dyes--Hoechst 33258, Chromomycin A3, and ethidium bromide--are reviewed. Fluorescence properties of these three stains when bound to mitotic cells or to chromosomes in suspension are measured and compared with fluorescence properties when bound to DNA in solution. Conditions are given for high resolution flow cytometry of Chinese hamster chromosomes stained with each of the fluorophors, and histograms are presented that exhibit differences in relative peak position and area. Energy transfer fluorescence between two DNA stains is presented as a potentially useful new parameter for flow cytometry of chromosomes and is illustrated by fluorescence energy transfer from Chromomycin A3 to ethidium bromide when simultaneously bound to hamster mitotic cells.

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DISTRIBUTION OF HETEROCHROMATIN IN THE CHROMOSOMES OF HAPLOPAPPUS GRACILIS

Canadian Journal of Genetics and Cytology ◽

10.1139/g68-058 ◽

1968 ◽

Vol 10 (2) ◽

pp. 433-443 ◽

Cited By ~ 6

Author(s):

Ira H. Ames ◽

Jyotirmay Mitra

Keyword(s):

Aqueous Solution ◽

Chromosome Aberrations ◽

Maleic Hydrazide ◽

Cold Treatment ◽

The Other ◽

Root Tip ◽

Metaphase Chromosomes ◽

Haplopappus Gracilis ◽

Tip Cells ◽

Root Tip Cells

Several approaches were employed to study the distribution of heterochromatin in root tip chromosomes of Haplopappus gracilis. Cold treatment and pretreatment in an aqueous solution of 8-hydroxyquinoline revealed achromatic gaps in metaphase chromosomes. Cold treatment also permitted the demonstration of positive heteropycnosis in prophase chromosomes. Further support for the identification of heterochromatic segments was provided by a study of the localization of chromosome aberrations induced by maleic hydrazide and an analysis of the pattern of DNA synthesis in chromosomes of root tip cells. Seven of the ten regions that were preferentially broken by maleic hydrazide also reacted differentially to cold treatment or to pretreatment with 8-hydroxyquinoline. A good correlation was found between regions that completed DNA replication late in the DNA-synnhetic period and segments that were shown to be heterochromatic by the other techniques.

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Flow Cytometry and Flow Sorting of Metaphase Chromosomes from the Dasyurid Marsupial Dasyurus viverrinus

Australian Journal of Biological Sciences ◽

10.1071/bi9850377 ◽

1985 ◽

Vol 38 (4) ◽

pp. 377 ◽

Cited By ~ 1

Author(s):

Brandon Wainwright ◽

Rory Hope

Keyword(s):

Flow Cytometry ◽

Chromosomal Dna ◽

Metaphase Chromosomes ◽

Homogeneous Groups ◽

Flow Sorting ◽

Dna Contents ◽

Conventional Methods

Metaphase chromosomes (2n = 14) from D. viverrinus were analysed by flow cytometry and flow sorted into six homogeneous groups. Relative chromosomal DNA contents and distribution frequencies of the groups corresponded closely with values for the karyotype obtained by conventional methods.

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Identification and Cytogenetic Analysis of an Abnormal Pig Chromosome for Flow Cytometry and Sorting

Zeitschrift für Naturforschung C ◽

10.1515/znc-1993-7-819 ◽

1993 ◽

Vol 48 (7-8) ◽

pp. 645-653 ◽

Cited By ~ 2

Author(s):

Michael Hausmann ◽

C. Paul Popescu ◽

Jeannine Boscher ◽

Dominique Kerboœf ◽

Jürgen Dölle ◽

...

Keyword(s):

Flow Cytometry ◽

Marker Chromosome ◽

Flow Analysis ◽

Chromosome 1 ◽

Cell Type ◽

Metaphase Chromosomes ◽

Flow Sorting ◽

Dna Libraries ◽

Sus Scrofa Domestica

Abstract For cytogenetics of pig (Sus scrofa domestica) and the influence of chromosome aberrations on pig production, high interest exists in flow sorted chromosomes for gene mapping, to establish DNA-libraries, or to produce DNA-probes. Flow karyotyping and sorting as well as slit scan flow analysis of metaphase chromosomes of an abnormal cell type carrying a translocation marker chromosome 6/15 are described. Flow sorting of the largest chromosomes of these cells was performed. After sorting the chromosomes still had a well preserved morphology and were identified microscopically by G-banding. The quality of the band pattern of the sorted chromosomes was compatible to that of isolated chromosomes not subjected to flow cytometry. The sorted fraction showed an enrichment of chromosom e 6/15 and chromosome 1 which have quantitatively about the same integrated fluorescence intensity. Slit scan flow analysis was performed to discriminate these two chromosomes. Metacentric and submetacentric chromosom es were analyzed according to their bimodal slit scan profiles. Profiles of the largest chromosomes were distinguished by their different centromeric indices. Two groups were interpreted as the normal chromosome 1 and the translocation chromosom e 6/15.

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