Cytochemical studies of metaphase chromosomes by flow cytometry

Chromosoma ◽  
1980 ◽  
Vol 77 (3) ◽  
pp. 229-251 ◽  
Author(s):  
R. G. Langlois ◽  
A. V. Carrano ◽  
J. W. Gray ◽  
M. A. Van Dilla
Keyword(s):  
Flow Cytometry ◽  
Metaphase Chromosomes

scholarly journals A template method for decomposing flow cytometry histograms of human chromosomes.

1979 ◽  
Vol 27 (1) ◽  
pp. 305-310 ◽  
Author(s):  
D H Moore
Keyword(s):  
Flow Cytometry ◽  
Trisomy 21 ◽  
Chromosomal Abnormalities ◽  
Simulated Data ◽  
Normal Karyotype ◽  
Template Method ◽  
Normal Person ◽  
Human Chromosomes ◽  
Metaphase Chromosomes ◽  
Flow Measurements

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.

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

Chromosoma ◽  
1984 ◽  
Vol 90 (4) ◽  
pp. 295-302 ◽  
Author(s):  
Barb Trask ◽  
Ger van den Engh ◽  
Joe Gray ◽  
Marty Vanderlaan ◽  
Bryan Turner
Keyword(s):  
Flow Cytometry ◽  
Metaphase Chromosomes

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

1977 ◽  
Vol 25 (8) ◽  
pp. 954-964 ◽  
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.

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

1985 ◽  
Vol 38 (4) ◽  
pp. 377 ◽  
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.

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

1993 ◽  
Vol 48 (7-8) ◽  
pp. 645-653 ◽  
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.

Some Structural Features of Metaphase Chromosomes of Mice and Men

1967 ◽  
Vol 25 ◽  
pp. 190-191
Author(s):  
Godfrey C. Hoskins ◽  
Betty B. Hoskins
Keyword(s):  
Electron Microscopy ◽  
Electron Micrographs ◽  
Structural Features ◽  
Metaphase Chromosomes ◽  
The Future ◽  
Of Mice And Men ◽  
Mouse Cells ◽  
Human And Mouse

Metaphase chromosomes from human and mouse cells in vitro are isolated by micrurgy, fixed, and placed on grids for electron microscopy. Interpretations of electron micrographs by current methods indicate the following structural features.Chromosomal spindle fibrils about 200Å thick form fascicles about 600Å thick, wrapped by dense spiraling fibrils (DSF) less than 100Å thick as they near the kinomere. Such a fascicle joins the future daughter kinomere of each metaphase chromatid with those of adjacent non-homologous chromatids to either side. Thus, four fascicles (SF, 1-4) attach to each metaphase kinomere (K). It is thought that fascicles extend from the kinomere poleward, fray out to let chromosomal fibrils act as traction fibrils against polar fibrils, then regroup to join the adjacent kinomere.

Studies of metaphase chromosomes in the scanning transmission x-ray microscope: Image fidelity, radiation damage and specimen preparation

1992 ◽  
Vol 50 (2) ◽  
pp. 1038-1039
Author(s):  
Shawn Williams ◽  
Xiaodong Zhang ◽  
Susan Lamm ◽  
Jack Van’t Hof
Keyword(s):  
Visible Light ◽  
Radiation Damage ◽  
Cross Section ◽  
Imaging Techniques ◽  
Dose Range ◽  
Specimen Preparation ◽  
X Rays ◽  
Metaphase Chromosomes ◽  
X Ray ◽  
Scanning Transmission

The Scanning Transmission X-ray Microscope (STXM) is well suited for investigating metaphase chromosome structure. The absorption cross-section of soft x-rays having energies between the carbon and oxygen K edges (284 - 531 eV) is 6 - 9.5 times greater for organic specimens than for water, which permits one to examine unstained, wet biological specimens with resolution superior to that attainable using visible light. The attenuation length of the x-rays is suitable for imaging micron thick specimens without sectioning. This large difference in cross-section yields good specimen contrast, so that fewer soft x-rays than electrons are required to image wet biological specimens at a given resolution. But most imaging techniques delivering better resolution than visible light produce radiation damage. Soft x-rays are known to be very effective in damaging biological specimens. The STXM is constructed to minimize specimen dose, but it is important to measure the actual damage induced as a function of dose in order to determine the dose range within which radiation damage does not compromise image quality.

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