R-banding of human chromosomes by heat denaturation and Giemsa staining after amethopterin-synchronization

1983 ◽  
Vol 25 (3) ◽  
pp. 261-269 ◽  
Author(s):  
C. L. Richer ◽  
M. Murer-Orlando ◽  
R. Drouin
Keyword(s):  
Giemsa Staining ◽  
Heat Denaturation ◽  
Human Chromosomes ◽  
Metaphase Chromosomes ◽  
Late Prophase ◽  
Chromosome Anomalies ◽  
Cell Synchronization

Human late prophase to late metaphase chromosomes were prepared after amethopterin cell synchronization. R-banding was produced by heat denaturation followed by Giemsa staining (RHG). Haploid sets of prophase chromosomes contain approximately 850 bands. Sequences of chromosomes of different degrees of condensation are presented; their analysis provides helpful information to identify the elongated chromosomes and to follow band subdivision. The heat denaturation technique is free from most of the disadvantages encountered with R-banding by 5-bromodeoxyuridine incorporation. Giemsa stained R-bands produced by heat denaturation on prophase and prometaphase chromosomes are useful to analyse the numerous chromosome anomalies involving R-bands. In conjunction with G-banding, it is also important to compare adequately the positive and negative regions of each chromosome to define the anomalies with precision.

Late replicating bands of human chromosomes demonstrated by fluorochrome and Giemsa staining

1975 ◽  
Vol 29 (1) ◽  
pp. 41-59 ◽  
Author(s):  
K. -H. Grzeschik ◽  
My. A. Kim ◽  
Renate Johannsmann
Keyword(s):  
Giemsa Staining ◽  
Human 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.

High-resolution idiogram of Giemsa R-banded human prophase chromosomes

1983 ◽  
Vol 25 (6) ◽  
pp. 642-650 ◽  
Author(s):  
C. L. Richer ◽  
R. Drouin ◽  
M. Murer-Orlando ◽  
P. Jean
Keyword(s):  
Comparative Analysis ◽  
High Resolution ◽  
Relative Position ◽  
Chromosomal Rearrangements ◽  
Staining Intensity ◽  
Chromosome Condensation ◽  
Giemsa Staining ◽  
Heat Denaturation ◽  
Schematic Representation ◽  
International Standard

The schematic representation of RHG-banded chromosomes (R-banding was produced by heat denaturation followed by Giemsa staining (RHG)) in the 850-band range per haploid set, was prepared showing the relative position, the specific size, and the characteristic staining intensity for each band. To this idiogram was adapted the new International Standard Cytogenetic Nomenclature. Our aim was to produce a realisitic idiogram which could help in the preparation of R-banded prophase karyotypes and in the localization of chromosomal rearrangements. A comparative analysis of bands at prophase and metaphase revealed certain aspects of the dynamics involved in chromosome condensation and in R-band organization. The effect of chromosome elongation on the appearance of R-bands within heterochromatic regions has also been discussed.

Analysis of high-resolution R-bands, obtained by heat-denaturation and Giemsa staining, on human prophase chromosomes

1985 ◽  
Vol 27 (1) ◽  
pp. 83-91 ◽  
Author(s):  
R. Drouin ◽  
C. L. Richer
Keyword(s):  
High Resolution ◽  
Banding Pattern ◽  
Giemsa Staining ◽  
Heat Denaturation ◽  
Banding Patterns ◽  
International Standard

RHG-bands (heat-denatured Giemsa R-bands) of human prophase chromosomes were analyzed at high resolution, and the banding patterns at prophase and metaphase are presented. The bands were compared with those of the International Standard Cytogenetic Nomenclature idiograms and of the G-band idiograms proposed by J. J. Yunis. The number, size, and position of the RHG-bands correspond rather well with their equivalent G-negative bands, but some differences were noted in the zones of preferential stretching, the juxtacentromeric regions, and the telomeres. Variations in the centromere index and the banding pattern in heterochromatin were also discussed.Key words: human prophase chromosomes, RHG-bands, high-resolution chromosomes.

C-banded karyotypes of Brassica campestris, B. oleracea, and B. napus

Genome ◽  
1992 ◽  
Vol 35 (4) ◽  
pp. 583-589 ◽  
Author(s):  
Majlis Olin-Fatih ◽  
W. K. Heneen
Keyword(s):  
Banding Pattern ◽  
Chromosome Pair ◽  
Brassica Campestris ◽  
Brassica Species ◽  
Conclusive Evidence ◽  
Metaphase Chromosomes ◽  
Late Prophase ◽  
Nucleolar Chromosome ◽  
Centromeric Position ◽  
Similar Banding Pattern

The chromosome complements of three Brassica species, namely B. campestris (2n = 20), B. oleracea (2n = 18), and B. napus (2n = 38), were studied using the air-dry method and C-banding. Karyotypes and ideograms of late prophase chromosomes were constructed, since contracted metaphase chromosomes were generally not suitable for this purpose. The three species generally had a similar banding pattern, manifested in the presence of a centromeric C-band in all chromosomes and heterochromatic knobs at the telomeric end of some chromosomes. The centromeric C-bands were more pronounced in B. campestris than in B. oleracea. Depending on the centromeric position, the chromosomes were grouped into median, submedian, subterminal, and terminal types. All chromosome pairs were morphologically distinguishable. Only one nucleolar chromosome pair, with heterochromatic satellites, was observed in each species. When compared, it was possible to distinguish chromosomes of both B. campestris and B. oleracea type in B. napus, but conclusive evidence as to the origin of all chromosome pairs in B. napus was not at hand.Key words: Brassica, chromosomes, late prophase, C-bands, knob structures, karyotypes, idiograms.

scholarly journals MICROFLUOROMETRIC ANALYSIS OF DEOXYRIBONUCLEIC ACID REPLICATION KINETICS AND SISTER CHROMATID EXCHANGES IN HUMAN CHROMOSOMES

1974 ◽  
Vol 22 (7) ◽  
pp. 478-491 ◽  
Author(s):  
SAMUEL A. LATT
Keyword(s):  
High Resolution ◽  
Dna Synthesis ◽  
Sister Chromatid ◽  
Deoxyribonucleic Acid ◽  
Sister Chromatid Exchanges ◽  
Human Chromosomes ◽  
Metaphase Chromosomes ◽  
Banding Patterns ◽  
Human Leukocytes ◽  
Chromatid Exchanges

Fluorescence of the dye 33258 Hoechst, when bound to chromosomes, is partially quenched by the incorporation of 5-bromodeoxyuridine into chromosomal deoxyribonucleic acid (DNA). This effect allows microfluorometric analysis of DNA synthesis. Metaphase chromosomes from cultured human leukocytes which have incorporated 5-bromodeoxyuridine for a portion of the DNA synthesis period exhibit reduced 33258 Hoechst fluorescence in 5-bromodeoxyuridine-containing regions. Regions synthesizing DNA during a particular interval can thus be highlighted by the appropriate protocol of 5-bromodeoxyuridine administration. Chromosomes from cells which have replicated twice in medium containing 5-bromodeoxyuridine exhibit one brightly and one dully fluorescing chromatid, reflecting incorporation of 5-bromodeoxyuridine into one or two chains of chromatid DNA, respectively. Sister chromatid exchanges, evident as sharply demarcated reciprocal alterations in fluorescence along chromosomes, can be located relative to quinacrine banding patterns. This fluorometric approach should be useful in many instances as a convenient, high resolution alternative to autoradiography.

scholarly journals Different sensitivity of DNA in situ in interphase and metaphase chromatin to heat denaturation.

1977 ◽  
Vol 73 (1) ◽  
pp. 128-138 ◽  
Author(s):  
Z Darzynkiewicz ◽  
F Traganos ◽  
T Sharpless ◽  
M R Melamed
Keyword(s):  
Heat Denaturation ◽  
Published Data ◽  
Dna Denaturation ◽  
Metaphase Chromosomes ◽  
Flow Cytofluorometry ◽  
Dna Reassociation ◽  
Interphase Cells ◽  
The Difference ◽  
The Stability

Heat denaturation of DNA in situ, in unbroken cells, was studied in relation to the cell cycle. DNA in metaphase cells denatured at lower temperatures (8 degrees-10 degrees C lower) than DNA in interphase cells. Among interphase cells, small differences between G1, S, and G2 cells were observed at temperatures above 90 degrees C. The difference between metaphase and interphase cells increased after short pretreatment with formaldehyde, decreased when cells were heated in the presence of 1 mM MgCl2, and was abolished by cell pretreatment with 0.5 N HCl. The results suggest that acid-soluble constituents of chromatin confer local stability to DNA and that the degree of stabilization is lower in metaphase chromosomes than in interphase nuclei. These in situ results remain in contrast to the published data showing no difference in DNA denaturation in chromatin isolated from interphase and metaphase cells. It is likely that factors exist which influence the stability of DNA in situ are associated with the super-structural organization of chromatin in intact nuclei and which are lost during chromatin isolation and solubilization. Since DNA denaturation is assayed after cell cooling, there is also a possibility that the extent of denatured DNA may be influenced by some factors that control strand separation and DNA reassociation. The different stainability of interphase vs. metaphase cells, based on the difference in stability of DNA, offers a method for determining mitotic indices by flow cytofluorometry, and a possible new parameter for sorting cells in metaphase.

BrdU-4Na-EDTA-Giemsa band karyotypes of 3 small freshwater fish, Danio rerio, Oryzias latipes, and Rhodeus ocellatus

Genome ◽  
1999 ◽  
Vol 42 (3) ◽  
pp. 531-535 ◽  
Author(s):  
T Ueda ◽  
H Naoi
Keyword(s):  
Freshwater Fish ◽  
Mitotic Index ◽  
Fish Species ◽  
Danio Rerio ◽  
Oryzias Latipes ◽  
Giemsa Staining ◽  
Banding Technique ◽  
Metaphase Chromosomes ◽  
Embryo Cells ◽  
Rhodeus Ocellatus

The 4Na-EDTA-Giemsa staining of metaphase chromosomes from embryos of three small freshwater fish, zebrafish Danio rerio, medakafish Oryzias latipes, and rosy bitterling Rhodeus ocellatus, in the presence of BrdU for one cycle gave rise to clear bands along the length of the chromosomes. These bands (B-bands) with G-band-like structures were clear and reproducible. However, as distinct B-bands were observed only in elongated chromosomes, fine chromosome preparations with a high mitotic index and elongated chromosomes were required. A technique for making preparations from embryo cells satisfied this request. The B-banding technique applied to embryo cells is useful to analyze chromosomes of fish species in which ordinary G-banding techniques have been known to bring about only unsatisfactory results.Key words: B-bands, karyotype, Danio rerio, Oryzias latipes, Rhodeus ocellatus.

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