scholarly journals DNA Measurement of Overlapping Cell Nuclei in Thick Tissue Sections

1997 ◽  
Vol 14 (1) ◽  
pp. 41-49 ◽  
Author(s):  
Liang Ji ◽  
James Tucker

The paper describes an improved image analysis procedure for measuring the DNA content of cell nuclei in thick sections of liver tissue by absorption densitometry. Whereas previous methods only permitted the analysis of isolated nuclei, the new technique enables both isolated and overlapping nuclei to be measured. A 3D segmentation procedure determines whether each object is an isolated nucleus or a pair of overlapping nuclei; in the latter case the combined optical density is redistributed to the individual nuclei. A selection procedure ensures that only complete nuclei are measured.The method has been tested on specially‐prepared Feulgen‐stained 20μsections of normal liver tissue. The overall distribution of the nuclear DNA measurements shows well‐defined diploid and tetraploid peaks, with coefficient of variations of less than 10%. Similar distributions were obtained from both the isolated nuclei and overlapped nuclei sub‐populations.

2011 ◽  
Vol 55 (1) ◽  
pp. 90-100 ◽  
Author(s):  
Christoph Krafft ◽  
Mehrnaz Alipour Diderhoshan ◽  
Peter Recknagel ◽  
Milos Miljkovic ◽  
Michael Bauer ◽  
...  

1997 ◽  
Vol 3 (S2) ◽  
pp. 1121-1122
Author(s):  
S.J. Lockett ◽  
E.G. Rodriguez ◽  
C. Ortiz de Solorzano ◽  
D. Sudar ◽  
D. Pinkel ◽  
...  

Combining pathology, which reports the structural features of individual cells and their spatial organi-zation in a tissue specimen, with molecular biology techniques (immunocytochemistry and fluores-cence in situ hybridization, FISH) for detecting the distribution of specific molecular species in individual cells is a powerful approach for gaining insight into the underlying disease mechanisms of carcinogenesis. This approach requires analysis of thick (>20 ¼m) tissue sections in which cells are preserved intact within the context of their environment. 3D (confocal) microscope image acquisition followed by 3D image analysis (IA) for extracting quantitative information are then used for quantita-tive analysis of tissue features such as nuclear and/or cell volume, shape, total fluorescence or FISH signal number. An essential component of the IA is detection of the individual cells, or their nuclei since many molecular species of interest are localized within the nucleus (e.g. genetic aberrations). We present here a completely automatic, 3D algorithm for this task, which based on the Hough transform (HT).


2004 ◽  
Vol 215 (1) ◽  
pp. 67-76 ◽  
Author(s):  
C. WAHLBY ◽  
I.-M. SINTORN ◽  
F. ERLANDSSON ◽  
G. BORGEFORS ◽  
E. BENGTSSON

1998 ◽  
Vol 21 (1) ◽  
pp. 47-54 ◽  
Author(s):  
Margarida Lima Carvalho ◽  
Claudio Oliveira ◽  
Fausto Foresti

The present paper reports nuclear DNA content in 30 Neotropical freshwater fish species and summarizes the data on other Neotropical species presented in the literature. Among Neotropical fishes, the nuclear DNA content ranges from 1.04 ± 0.09 pg/nucleus in Corydoras cf. simulatus (2n = 62) to 248.0 pg/nucleus in Lepidosiren paradoxa (2n = 38). A general analysis of the data obtained in the present study for each species showed that DNA measurements were practically constant at the individual level, while significant differences were observed among individuals of the same population. This observation was valid for all species analyzed and was more evident in those species that presented other karyotypic particularities such as sex chromosomes or supernumerary chromosomes. The importance of changes in nuclear DNA content in the evolutionary process of Neotropical fishes is discussed.


1985 ◽  
Vol 5 (3) ◽  
pp. 251-265 ◽  
Author(s):  
Ellen Spiesmacher ◽  
Hans-Peter Mühlbach ◽  
Martin Tabler ◽  
Heinz L. Sänger

Transcription studies with highly purified potato cell nuclei in combination with a ‘transcription-hybridization analysis’ unequivocally demonstrate that the nucleus is the subcellular site where the entire process of PSTV replication takes place. Inhibition experiments with actinomycin D and α-amanitin furthermore suggest that the nuclear DNA-dependent RNA polymerases I and II are involved in the synthesis of PSTV (+) and (−) RNA, respectively.


1978 ◽  
Vol 173 (1) ◽  
pp. 309-314 ◽  
Author(s):  
T R Butt ◽  
W M Wood ◽  
E L McKay ◽  
R L P Adams

The effects on DNA synthesis in vitro in mouse L929-cell nuclei of differential extraction of DNA polymerases alpha and beta were studied. Removal of all measurable DNA polymerase alpha and 20% of DNA polymerase beta leads to a 40% fall in the replicative DNA synthesis. Removal of 70% of DNA polymerase beta inhibits replicative synthesis by 80%. In all cases the nuclear DNA synthesis is sensitive to N-ethylmaleimide and aCTP (arabinosylcytosine triphosphate), though less so than DNA polymerase alpha. Addition of deoxyribonuclease I to the nuclear incubation leads to synthesis of high-molecular-weight DNA in a repair reaction. This occurs equally in nuclei from non-growing or S-phase cells. The former nuclei lack DNA polymerase alpha and the reaction reflects the sensitivity of DNA polymerase beta to inhibiton by N-ethylmaleimide and aCTP.


1974 ◽  
Vol 63 (2) ◽  
pp. 665-674 ◽  
Author(s):  
V. Mareš ◽  
B. Schultze ◽  
W. Maurer

Neurons of the mouse were labeled with [3H]thymidine during their prenatal period of proliferation. The 3H activity of the Purkinje cell nuclei was then studied autoradiographically 8, 25, 55, and 90 days after birth. The measured grain number per nucleus decreased by about 14% between the 8th and 25th postnatal days and then remained constant up to 90 days. There was no significant decrease of the 3H activity of the Purkinje cell nuclei after correction of the measured grain number per nucleus for increasing nuclear volume of the growing Purkinje cells and for the influence of [3H]ß self-absorption in the material of the sections. Injection of a high dose of [3H]thymidine into young adult mice did not result in 3H labeling of either Purkinje or other neurons in other brain regions. The results agree with the concept of metabolic stability of nuclear DNA. "Metabolic" DNA could not be observed in these experiments.


Cancers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 3108
Author(s):  
Jens Kleesiek ◽  
Benedikt Kersjes ◽  
Kai Ueltzhöffer ◽  
Jacob M. Murray ◽  
Carsten Rother ◽  
...  

Modern generative deep learning (DL) architectures allow for unsupervised learning of latent representations that can be exploited in several downstream tasks. Within the field of oncological medical imaging, we term these latent representations “digital tumor signatures” and hypothesize that they can be used, in analogy to radiomics features, to differentiate between lesions and normal liver tissue. Moreover, we conjecture that they can be used for the generation of synthetic data, specifically for the artificial insertion and removal of liver tumor lesions at user-defined spatial locations in CT images. Our approach utilizes an implicit autoencoder, an unsupervised model architecture that combines an autoencoder and two generative adversarial network (GAN)-like components. The model was trained on liver patches from 25 or 57 inhouse abdominal CT scans, depending on the experiment, demonstrating that only minimal data is required for synthetic image generation. The model was evaluated on a publicly available data set of 131 scans. We show that a PCA embedding of the latent representation captures the structure of the data, providing the foundation for the targeted insertion and removal of tumor lesions. To assess the quality of the synthetic images, we conducted two experiments with five radiologists. For experiment 1, only one rater and the ensemble-rater were marginally above the chance level in distinguishing real from synthetic data. For the second experiment, no rater was above the chance level. To illustrate that the “digital signatures” can also be used to differentiate lesion from normal tissue, we employed several machine learning methods. The best performing method, a LinearSVM, obtained 95% (97%) accuracy, 94% (95%) sensitivity, and 97% (99%) specificity, depending on if all data or only normal appearing patches were used for training of the implicit autoencoder. Overall, we demonstrate that the proposed unsupervised learning paradigm can be utilized for the removal and insertion of liver lesions at user defined spatial locations and that the digital signatures can be used to discriminate between lesions and normal liver tissue in abdominal CT scans.


1986 ◽  
Vol 83 (1) ◽  
pp. 77-87 ◽  
Author(s):  
M.D. Kendall ◽  
A. Warley

Mast cell granules were examined by fully quantitative X-ray microanalysis of 20 cells in freeze-dried cryosections. The mast cells were situated mainly in the connective tissue of the thymic capsule of five adult male Carworth Sprague Europe rats. In addition 30 red blood cells were analysed from the same sections. Nineteen of the mast cells had granules rich in S and K. One cell had smaller granules, and in this cell the granules contained high [Ca] and [P] instead of high [S] and [K]. In the majority of cells (13) the S:K ratio was highly correlated and less than 2.2, whereas in the remaining six cells the individual granule ratios were very variable in any one cell and much higher. The mean granule [K] (994 +/− 57 mmol kg-1 dry wt) was about four times the mean cytoplasmic level of 227 +/− 81 mmol kg-1 dry wt. The existence of this difference in concentration between the granules and the cytoplasm suggests that the K in the granules must be bound. The relationship between the [K] and [S] is discussed with regard to the possible binding of heparin and amines in the granules.


2015 ◽  
Vol 8 (5) ◽  
pp. 619-627 ◽  
Author(s):  
C. Pietsch ◽  
P. Burkhardt-Holm

Deoxynivalenol (DON) is a frequent contaminant of feeds in aquaculture, but the consequences of this contamination have rarely been evaluated. Previous studies on carp indicated effects of DON on liver function and histology after four weeks of feeding. The present study aimed to unravel the time course of liver responses of carp to orally applied DON. Therefore, liver enzyme activities and histology have been investigated after 7, 14, 26 and 56 days of DON feeding. The acute response comprises down-regulation of biotransformation enzymes, whereas the chronic response to DON is characterised by activation of alanine aminotransferase which indicates damage to liver tissue. Examination of histological sections of liver tissue revealed that changes such as fat aggregation, vacuolisation and hyperaemia were present after 14 and 26 days of exposure to DON but not thereafter. Several enzymes involved in glutathione cycling and reduction of oxidative stress were found to be reduced after 26 and 56 days of DON feeding. The results suggest that supporting the antioxidative system, e.g. by using glutathione-enriched yeast extracts as a food additive, might be successful in preventing the effects of DON in carp. This is the basis of a fundamental hypothesis since DON contamination of fish feed leads to pronounced effects on liver histology and liver enzyme activities which may also cause changes in the normal liver metabolism of endogenous and xenobiotic compounds.


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