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.

scholarly journals Purification of the chromosomes of the Indian muntjac by flow sorting.

1976 ◽  
Vol 24 (1) ◽  
pp. 348-354 ◽  
Author(s):  
A V Carrano ◽  
J W Gray ◽  
D H Moore ◽  
J L Minkler ◽  
B H Mayall ◽  
...  
Keyword(s):  
Large Fraction ◽  
Centromere Region ◽  
Chromosomal Dna ◽  
Metaphase Chromosomes ◽  
Banding Patterns ◽  
Indian Muntjac ◽  
Flow Sorting ◽  
Flow Microfluorometry ◽  
Mechanical Shearing ◽  
High Degree

Metaphase chromosomes were isolated from a male Indian muntjac cell line, were stained with ethidium bromide and were analyzed by flow microfluorometry to establish a deoxyribonucleic acid (DNA)-based karyotype. Five major peaks were evident on the chromosomal DNA distribution corresponding to the five chromosome types in this species. The amount of DNA in each chromosome was confirmed by cytophotometric measurements of intact metaphase spreads. The five chromosome types were separated by flow sorting at rates up to several hundred chromosomes per second. The sorted chromosomes were identified by morphology and by Giemsa banding patterns. The automsomes, Numbers 1, 2 and 3, and the X + 3 composite chromosome were separated with a high degree of purity (90%). The centromere region of the X + 3 chromosome was fragile to mechanical shearing, and during isolation a small proportion of these chromosomes broke into four segiments: the long arm, the short arm, the short arm plus centromere and the centromere region. A large fraction of the constitutive heterochromatin of this species is present in the centromere region of the X + 3 chromosome and in the Y chromosome; these two regions possess similar amounts of DNA and therefore sort together. Chromosome flow sorting is rapid, reproducible and precise; it allows the collection of microgram quantities of purified chromosomes.

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.

Image cytometric determination of nuclear and chromosomal DNA contents in Allium cepa L.

The Nucleus ◽  
2011 ◽  
Vol 54 (2) ◽  
pp. 71-75
Author(s):  
V. Rao Bandaru ◽  
Tavva S. S. Mohan Dev ◽  
P. V. Arjuna Rao ◽  
M. V. Subba Rao
Keyword(s):  
Allium Cepa ◽  
Chromosomal Dna ◽  
Allium Cepa L ◽  
Dna Contents

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.

scholarly journals Electron microscopy of gold-labeled human and equine chromosomes.

1989 ◽  
Vol 37 (9) ◽  
pp. 1443-1447 ◽  
Author(s):  
P E Messier ◽  
R Drouin ◽  
C L Richer
Keyword(s):  
Electron Microscopy ◽  
Chromosome Banding ◽  
Protein A ◽  
Gold Complex ◽  
Chromosome Identification ◽  
Chromosomal Dna ◽  
Metaphase Chromosomes ◽  
Sister Chromatids ◽  
Antigen Localization ◽  
Chromatin Reorganization

We present an immunochemical technique for the detection of 5-bromo-2'-deoxyuridine (BrdU) incorporated discontinuously into the chromosomal DNA. A monoclonal anti-BrdU antibody and a protein A-gold complex were used to produce chromosome banding of human and equine chromosomes, specific for electron microscopy (EM). Well-defined bands, symmetry of sister chromatids, concordance between homologues, and band patterns similar to those observed by light microscopy facilitate chromosome identification and karyotyping. From prophase to late metaphase, chromosomes condense and bands appear to fuse. The fusion appears to be owing to chromatin reorganization. Our results underline the value of using immunogold reagents, which are ideal probes for antigen localization on chromosomes.

scholarly journals Measurement of transcribed human X-chromosomal DNA sequences transferred to rodent cells by chromosome-mediated gene transfer.

1982 ◽  
Vol 2 (1) ◽  
pp. 52-65 ◽  
Author(s):  
O W McBride ◽  
A S Olsen ◽  
G S Aulakh ◽  
R S Athwal
Keyword(s):  
Dna Sequences ◽  
Genetic Information ◽  
Hybrid Cell ◽  
Single Copy ◽  
Nucleic Acid Hybridization ◽  
Chromosomal Dna ◽  
Metaphase Chromosomes ◽  
Transfer Event ◽  
Donor Chromosome ◽  
Labeled Probe

Transfer of genetic information can be effected by incubation of cultured eucaryotic cells with isolated metaphase chromosomes. In most cases, a resulting transformed cell contains only a fragment of a donor chromosome. The amount of transferred donor DNA has been quantified in 11 independent mouse A9 transformants by nucleic acid hybridization analysis. Each transformant had been selected for hprt (hypoxanthine phosphoribosyltransferase; EC 2.4.2.8) transfer and contained part of the human X chromosome. A labeled probe of transcribed human X-chromosomal DNA was prepared by hybridization of nick-translated unique-sequence human DNA with whole cellular RNA from a human-mouse hybrid cell line, A9/HRBC2-A, containing a single human chromosome., X. The amount of human X-chromosomal DNA in the transformants was quantitated by comparing the hybridization of this probe with transformant and A9/HRBC2-A DNAs. Two unstable transformants which had a microscopically detectable donor chromosome fragment contained 15% of the human X-chromosomal single-copy DNA. Four other unstable transformants contained 4 to 7% of human X-chromosomal DNA sequences. The transferred DNA was below the level of detection in three other unstable and in all three stable transformants. We conclude that the initial transfer event can introduce a substantial amount of genetic information but only smaller amounts of DNA are stably incorporated by integration.

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