Atomic force microscope imaging of chromosome structure during G-banding treatments

Musio, Antonio; Mariani, Tullio; Frediani, C.; Ascoli, C.; Sbrana, Isabella

doi:10.1139/g97-018

Cited by 29 publications

(24 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A similar behavior could be due to the protein precipitating capabilities of the detergent solution (Miller et al 1988). Moreover, comparable features have also been detected during different steps of the Giemsa chromosome staining by AFM (Musio et al 1997). In spite of these large round aggregates, the 10 nm fibers chromatin network, detectable in clean areas of the samples by high-magnification FEISEM analysis, is completely comparable with the network structure that can be seen accurately on untreated chromosomes selected without coating (Rizzoli et al 1994).…”

Section: Discussionsupporting

confidence: 68%

“…It is well known that many banding methods can be used to identify individual chromosomes (i.e., the G-banding) but, although these techniques are used routinely for conventional cytogenetic investigations, many of the mechanisms involved are not completely understood (Musio et al 1997). A longitudinal pattern similar to the G-banding can be clearly detected by AFM on the chromosome surface after adequate aging of the maps, or during the treatments required to perform the G-banding that involves a proteolytic digestion of the sample (Ali Ergun et al 1999;Mariani et al 1994;Musio et al 1994Musio et al , 1997.…”

Section: Discussionmentioning

confidence: 99%

“…A longitudinal pattern similar to the G-banding can be clearly detected by AFM on the chromosome surface after adequate aging of the maps, or during the treatments required to perform the G-banding that involves a proteolytic digestion of the sample (Ali Ergun et al 1999;Mariani et al 1994;Musio et al 1994Musio et al , 1997. Under our conditions, samples were dehydrated and dried shortly after the spontaneous air drying of the methanol-acetic acid fixative: a G-banding-like pattern is not detectable, except for the chromosomes treated with the solution with Na-acetate, protease K, and SDS and rehydrated; however, these chromosomes are not completely coat-free.…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Untitled

2000

Scanning

View full text Add to dashboard Cite

Summary:A correlative morphologic analysis was carried out on isolated metaphase chromosomes by means of field emission in-lens scanning electron microscopy (FEISEM) and atomic force microscopy (AFM). Whereas FEISEM provides ultra-high resolution power and allows the surface analysis of biological structures free of any conductive coating, the AFM allows imaging of biological specimens in ambient as well as in physiologic conditions. The analysis of the same samples was made possible by the use of electrical conductive and light transparent ITO glass as specimen holder. Further preparation of the specimen specific for the instrumentation was not required. Both techniques show a high correlation of the respective morphologic information, improving their reciprocal biological significance. In particular, the biological coat represents a barrier for surface morphologic analysis of chromosome spreads and it is sensitive to protease treatment. The chemical removal of this layer permits high-resolution imaging of the chromatid fibers but at the same time alters the chromosomal dimension after rehydration. The high-resolution level, necessary to obtain a precise physical mapping of the genome that the new instruments such as FEISEM and AFM could offer, requires homogeneously cleaned samples with a high grade of reproducibility. A correlative microscopical approach that utilizes completely different physical probes provides complementary useful information for the understanding of the biological, chemical, and physical characteristics of the samples and can be applied to optimize the chromosome preparations for further improvement of the knowledge about spatial genome organization.

show abstract

Section: Discussionsupporting

confidence: 68%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Untitled

2000

Scanning

View full text Add to dashboard Cite

show abstract

“…Atomic force microscopy as a new diagnostic tool for further chromosomal studies allows detection of the de- tailed structure of human chromosomes (Musio et al 1997).…”

Section: Discussionmentioning

confidence: 99%

“…Atomic force microscopy (AFM) is a diagnostic tool for detecting detailed structures of the chromosomes, and the surface topography of human chromosomes can be examined using this new technique (Binnig et al 1986, Musio et al 1997. The images of AFM are compatible with previous scanning electron microscopy studies (Harrison et al 1983).…”

Section: Introductionmentioning

confidence: 99%

Determination of a translocation chromosome by atomic force microscopy

et al. 2002

View full text Add to dashboard Cite

Summary: Atomic force microscopy (AFM) has been used to study the translocation involving chromosomes 11 and 13. An amniocentesis procedure was performed at 18 weeks of pregnancy on a familial balanced translocation carrier mother whose karyotype was 46,XX,t(11;13) (q23;q34). After harvesting the tissue cultures, light microscopy studies (LM) have indicated that the fetus had the same translocation. A 0.3 µm gap region on the derivative chromosome 13 was determined by AFM; it was equivalent to a mid-sized G-band. The enhanced resolution of AFM with respect to its line measure analysis and threedimensional image capture capability has allowed an extension and reconsideration of conclusions about chromosomal aberrations based on the study of LM preparations. In this manner, chromosomal disorders will be studied at nanoscale to help in the planning of new therapy strategies.

show abstract

Cytogenetic stability of chicken T‐cell line transformed with Marek's disease virus: atomic force microscope, a new tool for investigation

Bucchianico¹,

Giardi²,

Marco³

et al. 2010

J of Molecular Recognition

View full text Add to dashboard Cite

The Marek's disease virus (MDV) integration may induce a novel organization of chromatin architecture with a modified genetic expression. In our opinion it is worthwhile trying to relate cytogenetic stability to functional modifications. Recently, atomic force microscopy technique was applied to study the structure of chromosomes at a nanoscale level. This high resolution allows to investigate the different structure of chromatin in order to study cytogenetic stability and chromosome aberrations due to MDV insertion. In this paper data are presented indicating a duplication [78,WZ,dup(1p)(p22-p23)] and a deletion [78,WZ cht del(3)(q2.10)] of Chromosomes 1 and 3 relatively. Relationships between GTG (G-bands by Trypsin using Giemsa) bands and the topography of chromosomes are also discussed, naming them Topographic Banding. The architecture of chromosomes observed by AFM can be related to the data obtained with classic banding techniques thus overcoming the optical resolution limits. The presence of chromatin bridges between sister chromatids at most of the heterochromatic regions is also evidenced. Besides, we present different studies of the longitudinal and transversal symmetry of the hetero and euchromatic regions to clearly demonstrate a different underlying architecture of these regions. It is indeed evident that the heterochromatic bands are more symmetrical than euchromatic bands.

show abstract

Atomic force microscope imaging of chromosome structure during G-banding treatments

Cited by 29 publications

References 14 publications

Untitled

Untitled

Determination of a translocation chromosome by atomic force microscopy

Cytogenetic stability of chicken T‐cell line transformed with Marek's disease virus: atomic force microscope, a new tool for investigation

Contact Info

Product

Resources

About