Fluorescence image cytometry for measurement of nuclear DNA content in surgical pathology

Cytometry ◽  
1995 ◽  
Vol 22 (4) ◽  
pp. 323-329 ◽  
Author(s):  
Naining Wang ◽  
Yi Pan ◽  
Thomas Heiden ◽  
Bernhard Tribukait
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Image Cytometry ◽  
Surgical Pathology ◽  
Fluorescence Image

scholarly journals Fluorescence image cytometry of nuclear DNA content versus chromatin pattern: a comparative study of ten fluorochromes.

1992 ◽  
Vol 40 (11) ◽  
pp. 1789-1797 ◽  
Author(s):  
M S Santisteban ◽  
M P Montmasson ◽  
F Giroud ◽  
X Ronot ◽  
G Brugal
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Image Cytometry ◽  
Binding Mode ◽  
Fluorescence Image ◽  
Chromatin Pattern ◽  
Fluorescent Image ◽  
Mouse Hepatocytes

This study is intended to be the first step of an in situ exploration of the intranuclear DNA distribution by image cytometry (SAMBA) with several fluorochromes. The nuclear DNA content and the chromatin pattern, revealed by ten fluorochromes (HO, DAPI, MA, CMA3, OM, QM, AO, EB, PI, and 7-AMD), were analyzed on mouse hepatocytes fixed by the Boehm-Sprenger procedure optimal for preserving the chromatin pattern. The question was whether fluorochromes specific to DNA make it possible to accurately quantitate the total nuclear DNA content when the chromatin pattern is preserved. Only HO and MA were found to provide satisfactory quantitation of nuclear DNA content, as assumed by both a small CV and a 4c to 2c ratio equal to 2. PI, EB, 7-AMD, and OM provided higher CV values, although the 4c to 2 c ratio was still equal to 2. QM, AO, CMA3, and DAPI provided non-reproducible and non-stoichiometric nuclear DNA content measurements under the fixation conditions used. The intranuclear and the internuclear SD of the fluorescence intensities describing the fluorescence pattern of the 2c hepatocytes proved to vary according to both the basepair specificity and the binding mode of the fluorochromes. The results reported here argue in favor of an external binding of 7-AMD to DNA and an increased quantum yield of QM when bound to AT-rich DNA. For PI, EB, 7-AMD, and OM, the measured DNA content increased with the fluorescence distribution heterogeneity. This correlation was not observed with other fluorochromes and is suggested to result from decreased fluorochrome accessibility to DNA when the chromatin is condensed. This study demonstrates that under conditions that preserve chromatin organization, only HO for AT-rich DNA and MA for GC-rich DNA can be used, alone or in combination, to measure nuclear DNA content. With other fluorochromes, either the measured DNA content or the chromatin pattern is assessed in suboptimal conditions when fluorescent image cytometry is used.

Nuclear DNA content of three Eucalyptus species estimated by flow and image cytometry

2009 ◽  
Vol 57 (6) ◽  
pp. 524 ◽  
Author(s):  
Milene Miranda Praça ◽  
Carlos Roberto Carvalho ◽  
Carolina Ribeiro Diniz Boaventura Novaes
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Methodological Approach ◽  
Nuclear Dna Content ◽  
Image Cytometry ◽  
Eucalyptus Species ◽  
International Consensus ◽  
Mean Values ◽  
Dna Contents ◽  
Nuclear Dna Contents

Previous flow cytometry (FCM) analyses delivered nearly equal mean values of nuclear 2C DNA content for Eucalyptus grandis Hill ex Maiden and E. urophylla S. T. Blake (1.33 pg and 1.34 pg, respectively), whereas E. globulus Labill. presented distinct mean values (1.09, 1.13 and 1.40). These differences have been attributed to the different methodological approach, utilised plant cultivar and presence of intrinsic metabolic compounds that affect fluorochrome fluorescence. In the present study, a FCM and image cytometry (ICM) design, following international consensus criteria, were adopted to reassess the nuclear DNA contents of the above-mentioned Eucalyptus species. Statistical analyses revealed either similar or discrepant nuclear DNA contents, depending on the standard species used and whether FCM or ICM was employed. Our results demonstrated that 2C DNA values obtained by FCM and ICM were most uniform when Solanum lycopersicum was used as a standard. Moreover, the values obtained for E. grandis and E. urophylla were close, but differed as much as 24.63% in relation to previous data, and E. globulus proportionally varied up to 25%. New DNA content values are suggested for these eucalypt species.

scholarly journals Nuclear DNA Content and Chromatin Pattern of Rat Rhabdomyosarcoma Cell Sublines with Different Metastatic Potentials

2000 ◽  
Vol 20 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Jean Dufer ◽  
Marie-France Poupon ◽  
Sonia Yatouji
Keyword(s):  
Tumor Cells ◽  
Cell Lines ◽  
Dna Content ◽  
Nuclear Dna ◽  
Chromatin Condensation ◽  
Nuclear Dna Content ◽  
Image Cytometry ◽  
Chromatin Pattern ◽  
Rhabdomyosarcoma Cell

There is a constant need of features able to characterize potentially metastatic cells among the heterogeneous cell subpopulations which constitute a tumor. Image cytometry of metastatic tumor cells give rise to variable results, partly because of a heterogeneous origin of cells, or potential drug effects. The aim of this work was to characterize nuclear changes observed in metastatic cell clones issuedin vitrofrom the same parental cell population The nuclear phenotypes of 6 cell sublines isolated from a rat rhabdomyosarcoma cell line and differing in their metastatic ability were evaluated by image cytometry on Feulgen‐stained preparations. Densitometric [5], geometric [3] and textural [9] features were computed from each nuclear image. For each cell subline, a metastatic score, ranging from 0 to 10, was calculated on the basis ofin vitroinvasivity data, by measuring the number of pulmonary metastases observed after s.c. graft of tumor cells in rats. Data obtained were compared to karyotype, growth characteristics, and oncogene expressions of cell lines. The nuclear DNA content, the chromosome numbers, the cell sublines doubling times, and the distribution of cells within the cell cycle appear unrelated with this score. On the contrary, increase in metastatic ability is accompanied by changes in chromatin pattern as assessed by textural features. Progressive increase in chromatin condensation can be observed in cell sublines with increasing metastatic score. These results were confirmed by an unsupervised multivariate partitioning of rhabdomyosarcoma cells which identified two separate subsets whose distributions within the analyzed cell lines correlate with their metastatic ability. These data suggest that, in rat rhabdomyosarcoma cell sublines, metastatic ability could be associated with nuclear morphological changes at the level of chromatin texture.

Prognostic value of nuclear DNA content in papillary and follicular thyroid cancer

1988 ◽  
Vol 12 (4) ◽  
pp. 503-507 ◽  
Author(s):  
Jaap F. Hamming ◽  
Lodewijk J. D. M. Schelfhout ◽  
Cees J. Cornelisse ◽  
Cornelis J. H. van de Velde ◽  
Bernard M. Goslings ◽  
...  
Keyword(s):  
Thyroid Cancer ◽  
Dna Content ◽  
Prognostic Value ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Follicular Thyroid Cancer

Estimation of nuclear DNA content by flow cytometry in fishes of the genus Xiphophorus

1989 ◽  
Vol 94 (3) ◽  
pp. 465-468 ◽  
Author(s):  
Terrence R. Tiersch ◽  
Robert W. Chandler ◽  
Klaus D. Kallman ◽  
Stephen S. Wachtel
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content
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