Determination of cellular DNA content of Iberian salamanders by flow cytometry

2000 ◽  
Vol 21 (4) ◽  
pp. 411-418 ◽  
Author(s):  
Miguel Lizana ◽  
Roberto Martín-Sánchez ◽  
Rafael Márquez ◽  
Juana Ciudad ◽  
Antonio López ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Reproductive Biology ◽  
Dna Content ◽  
Diploid Nucleus ◽  
C Value ◽  
Salamandra Salamandra ◽  
Cellular Dna ◽  
Pleurodeles Waltl

AbstractThe DNA content per diploid nucleus (2 C-value) was determined for the eight species of Iberian salamanders by flow cytometry. All species showed high values, which are characteristic of Caudata. The species with the lowest value was Pleurodeles waltl (48.25 pg DNA/cell) and Salamandra salamandra had the highest values (70.55 pg DNA/cell). There were no statistical differences between sexes in any of the eight species and DNA content was not directly related with phylogeny. The values obtained are often higher than previously published estimates obtained with older and potentially less accurate methodologies. We discuss the possible relationship between cellular DNA content and reproductive biology. Se determinó el contenido en ADN por núcleo diploide (valor 2-C) por citometría de flujo de ocho especies de salamandras ibéricas. Todas las especies mostraron altos valores, lo que es característico de Caudata. La especie con el menor valor fue Pleurodeles waltl (48,25 pg ADN/célula) mientras Salamandra salamandra tuvo las valores más altos (70,55 pg ADN/célula). No se encontraron diferencias estadísticas entre sexos en ninguna de las 8 especies y el contenido en ADN no se relacionó directamente con la filogenia. Los valores obtenidos son, a menudo, más altos que los publicados previamente con técnicas más antigüas, y potencialmente menos precisas. Discutimos las posibles relaciones entre el contenido en ADN celular y la biología reproductiva de cada especie.

Flow cytometry, cellular DNA content, and prognosis in human malignancy.

1987 ◽  
Vol 5 (10) ◽  
pp. 1690-1703 ◽  
Author(s):  
D E Merkel ◽  
L G Dressler ◽  
W L McGuire
Keyword(s):  
Flow Cytometry ◽  
Data Base ◽  
Dna Content ◽  
Dna Analysis ◽  
Clinical Decision Making ◽  
Risk Groups ◽  
Clinical Decision ◽  
Tumor Type ◽  
Cellular Dna ◽  
Bladder Carcinomas

The use of flow cytometry to analyze the cellular DNA content of human malignancies has become increasingly commonplace. The relationship between abnormalities in DNA content or proliferative characteristics and prognosis is becoming clear for a variety of malignancies in part through new techniques that permit analysis of archival material. High- and low-risk groups of patients with early breast and bladder carcinomas, non-small-cell lung cancer, and colorectal, ovarian, and cervical carcinoma can be distinguished on the basis of abnormal stemline DNA content. In several hematologic and common pediatric malignancies, the prognostic relevance of DNA content flow cytometry has been similarly established. Though the interpretation of tumor cell cycle analyses is less certain, this characteristic may also be prognostically important. However, generalizations cannot be made when applying flow cytometric DNA analysis to clinical decision making. The prognostic importance of an abnormal DNA histogram for an individual patient must be assessed on the basis of the relevant data base for that particular tumor type. The current extent of this data base for various malignancies is reviewed.

Prognostic Value of Flow Cytometry in Bladder Tumours

1993 ◽  
Vol 60 (2) ◽  
pp. 152-157
Author(s):  
D. Grassi ◽  
M. De Siati ◽  
N. Franzolin
Keyword(s):  
Flow Cytometry ◽  
Transitional Cell Carcinoma ◽  
Cell Carcinoma ◽  
Dna Content ◽  
Prognostic Value ◽  
Transitional Cell ◽  
Prognostic Information ◽  
Carcinoma Of The Bladder ◽  
Bladder Tumours

During this last decade, flow cytometry (FCM) has been widely investigated and employed in assessing the DNA content of bladder tumours. The prognostic value of FCM is recognised by the majority of investigators, above all when it concerns superficial transitional cell carcinoma of the bladder. The determination of ploidy and the degree of aneuploidy seem to offer valuable prognostic information. The Authors have reviewed Literature on this subject, identifying three different categories of studies that analyse the relation of FCM to cytohystological characterisation, to the clinical behaviour of the tumours and to the patients’ survival.

scholarly journals Method for analysis of cellular DNA content of paraffin-embedded pathological material using flow cytometry.

1983 ◽  
Vol 31 (11) ◽  
pp. 1333-1335 ◽  
Author(s):  
D W Hedley ◽  
M L Friedlander ◽  
I W Taylor ◽  
C A Rugg ◽  
E A Musgrove
Keyword(s):  
Flow Cytometry ◽  
Retrospective Study ◽  
Clinical Outcome ◽  
Dna Content ◽  
Prognostic Significance ◽  
Distilled Water ◽  
Archival Material ◽  
Cellular Dna ◽  
Pathological Material ◽  
Dna Histograms

A method has been developed that allows flow cytometry to be used for measuring the cellular DNA content of paraffin-embedded human tumors. Thick (i.e., 30 micron) sections were cut from tissue blocks using a microtome and dewaxed in xylene. The sections were then rehydrated by sequentially immersing them in 100, 95, 70, and 50% ethanol before finally washing in distilled water. Single cell suspensions were then prepared by incubation in 0.5% pepsin, pH 1.5, at 37 degrees C for 30 min. The cells were counted, washed, and stained with 1 microgram/ml 4',6'-diamidino-2-phenylindole for 30 min, and DNA content was measured using an ICP 22 flow cytometer. There was a good correlation between the DNA histograms produced using this method and those obtained using unfixed tissue from the same tumor stained with ethidium bromide plus mithramycin. This method allows the retrospective study of archival material where the clinical outcome is already known, and it should, therefore, be particularly useful for determining the prognostic significance of abnormal DNA content measured by flow cytometry.

scholarly journals NUCLEAR DNA CONTENT OF DENDROBIUM ORCHID SPECIES AS DETERMINED BY LASER FLOW CYTOMETRY

HortScience ◽  
1996 ◽  
Vol 31 (3) ◽  
pp. 322c-322
Author(s):  
W.E. Jones ◽  
A.R. Kuehnle ◽  
K. Arumuganathan
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Nuclear Dna ◽  
Evolutionary Relationship ◽  
Internal Standard ◽  
Nuclear Dna Content ◽  
Orchid Species ◽  
Group Assignment ◽  
Taxonomic Groups

Flow cytometry (FC) has proven to be an efficient and reliable method to estimate nuclear DNA content (genome size) in quantifiable units useful for genetic and molecular biology studies. This method also makes possible determination of the variation in nuclear DNA content between related taxa, which gives insights into the process of speciation. In this study, DNA content was determined in nuclei isolated from leaves of 21 Dendrobium species representing each of the major taxonomic groups used in the Univ. of Hawaii breeding program. Nuclei were mechanically isolated, stained with the nucleic acid-specific fluorochrom propidium iodide, and DNA content determined using a Coulter Epics 753 laser flow cytometer. Chicken erythrocyte nuclei (2C = 2.33 pg DNA) were used as an internal standard for direct comparative measurement. The mean diploid genome (2C) values for Dendrobium species ranged from 3.36 to 5.06 pg. Genome sizes were evaluated for possible use as discrete characters for taxonomic group assignment and compared to previous data on breeding compatibility and evolutionary relationship between species.

scholarly journals The coexistence of acute myeloblastic leukemia and diffuse histiocytic lymphoma in the same patient as demonstrated by multiparameter analysis

Blood ◽  
1979 ◽  
Vol 54 (6) ◽  
pp. 1428-1433
Author(s):  
DJ Straus ◽  
M Andreeff ◽  
HJ Hansen ◽  
R Mertelsmann ◽  
B Koziner ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Lymph Node ◽  
Dna Content ◽  
Acute Myeloblastic Leukemia ◽  
Histiocytic Lymphoma ◽  
Cellular Dna ◽  
Multiparameter Analysis ◽  
Diploid Cells ◽  
Diffuse Histiocytic Lymphoma

Measurement of cellular DNA content by flow cytometry demonstrated presence of two distinct aneuploid neoplasms in a patient who developed acute myeloblastic leukemia (AML) 4 mo after diagnosis of a diffuse histiocytic lymphoma (DHL). A lymph node aspirate contained peroxidase- negative, “null,” hyperdiploid (2.6C) DHL cells, while the bone marrow (BM) contained 84% primitive peroxidase-positive tetraploid AML cells (4.0C). Minor populations of hyperdiploid HDL and normal diploid cells could be detected by flow-cytometry in the BM, and all three populations were also seen in the peripheral blood.

Analysis of Cellular DNA Content by Flow Cytometry

2017 ◽  
Vol 82 (1) ◽  
Author(s):  
Zbigniew Darzynkiewicz ◽  
Xuan Huang ◽  
Hong Zhao
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Cellular Dna

scholarly journals Measurement of cellular DNA mass by flow microfluorometry with use of a biological internal standard.

1978 ◽  
Vol 26 (2) ◽  
pp. 145-148 ◽  
Author(s):  
E Tannenbaum ◽  
M Cassidy ◽  
O Alabaster ◽  
C Herman
Keyword(s):  
Dna Content ◽  
Internal Standard ◽  
Cell Populations ◽  
Experimental Cell ◽  
Flow Microfluorometry ◽  
Chicken Erythrocytes ◽  
Cellular Dna ◽  
Biological Standard

Use of a biological standard (chicken erythrocytes) mixed with experimental cell populations allows control of all variables in flow microfluorometric determination of DNA content. These variables include both staining and instrument procedures. In addition, the use of a biological standard allows determination of cellular DNA mass in unperturbed cell populations. DNA mass measured by FMF technique correlates closely with values reported in the literature that used biochemical techniques.

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