• Title/Summary/Keyword: Image Cytometry

Search Result 20, Processing Time 0.033 seconds

DNA Ploidy in Anaplastic Carcinoma of the Thyroid Gland by Image Analysis (갑상선 역형성암종의 DNA 배수성에 관한 화상분석학적 연구)

  • Lee, Ji-Shin;Lee, Min-Cheol;Park, Chang-Soo;Juhng, Sang-Woo
    • The Korean Journal of Cytopathology
    • /
    • v.6 no.1
    • /
    • pp.10-17
    • /
    • 1995
  • Anaplastic carcinoma of the thyroid gland is one of the most malignant tumors. Recently, DNA ploidy measured by flow cytometry and image analysis has been suggested as an additional useful indicator of tumor behavior. Studies on the occurrence and clinical significance of DNA aneuploidy in anaplastic carcinoma of the thyroid are rare. In this study, the pattern of DNA ploidy was measured by image analysis on Papanicolaou stained slides in four cases of anaplastic carcinoma and also measured by flow cytometry using paraffin blocks in two cases. In all cases of anaplastic carcinoma, DNA aneuploidy was found by image analaysis. By flow cytometry, one case had a diploid peak and the other case had an aneuploid peak. According to the above results, we conclude that anaplastic carcinoma of the thyroid glands have a high incidence of DNA aneuploidy and image analysis using Papanicolaou stained slides is a useful method in detecting DNA aneuploidy.

  • PDF

Microfluidic Image Cytometry (μFIC) Assessments of Silver Nanoparticle Cytotoxicity

  • Park, Jonghoon;Yoon, Tae Hyun
    • Bulletin of the Korean Chemical Society
    • /
    • v.33 no.12
    • /
    • pp.4023-4027
    • /
    • 2012
  • Cytotoxicity assessment of silver nanoparticles (AgNPs) was performed using MTT-based microfluidic image cytometry (${\mu}FIC$). The $LC_{50}$ value of HeLa cells exposed to AgNPs in the microfluidic device was estimated as 46.7 mg/L, which is similar to that estimated by MTT-based IC for cells cultured in a 96 well plate (49.9 mg/L). These results confirm that the ${\mu}FIC$ approach can produce cytotoxicity data that is reasonably well-matched with that of the conventional 96 well plate system with much higher efficiency. This ${\mu}FIC$ method provides many benefits including ease of use and low cost, and is a more rapid in vitro cell based assay for AgNPs. This may aid in speeding up data acquisition in the field of nanosafety and make a significant contribution to the quantitative understanding of nanoproperty-toxicity relationships.

Effects of Ag Nanoparticle Flow Rates on the Progress of the Cell Cycle Under Continuously Flowing "Dynamic" Exposure Conditions

  • Park, Min Sun;Yoon, Tae Hyun
    • Bulletin of the Korean Chemical Society
    • /
    • v.35 no.1
    • /
    • pp.123-128
    • /
    • 2014
  • In this study, we have investigated the flow rate effects of Ag nanoparticle (NP) suspensions on the progress of the cell cycle by using a microfluidic image cytometry (${\mu}FIC$)-based approach. Compared with the conventional "static" exposure conditions, enhancements in G2 phase arrest were observed for the cells under continuously flowing "dynamic" exposure conditions. The "dynamic" exposure conditions, which mimic in vivo systems, induced an enhanced cytotoxicity by accelerating G2 phase arrest and subsequent apoptosis processes. Moreover, we have also shown that the increases in delivered NP dose due to the continuous supply of Ag NPs contributed dominantly to the enhanced cytotoxicity observed under the "dynamic" exposure conditions, while the shear stress caused by these slowly flowing fluids (i.e., flow rates of 6 and $30{\mu}L/h$) had only a minor influence on the observed enhancement in cytotoxicity.

Lamin A/C and Polymeric Actin in Genome Organization

  • Ondrej, Vladan;Lukasova, Emilie;Krejci, Jana;Matula, Pavel;Kozubek, Stanislav
    • Molecules and Cells
    • /
    • v.26 no.4
    • /
    • pp.356-361
    • /
    • 2008
  • In this work, we have studied the structural and functional linkage between lamin A/C, nuclear actin, and organization of chromosome territories (CTs) in mammary carcinoma MCF-7 cells. Selective down-regulation of lamin A/C expression led to disruption of the lamin A/C perinuclear layer and disorganization of lamin-bound emerin complexes at the inner nuclear membrane. The silencing of lamin A/C expression resulted in a decrease in the volume and surface area of chromosome territories, especially in chromosomes with high heterochromatin content. Inhibition of actin polymerization led to relaxation of the structure of chromosome territories, and an increase in the volumes and surface areas of the chromosome territories of human chromosomes 1, 2 and 13. The results show an important role of polymeric actin in the organization of the nuclei and the chromosome territories.

Use of DNA-Specific Anthraquinone Dyes to Directly Reveal Cytoplasmic and Nuclear Boundaries in Live and Fixed Cells

  • Edward, Roy
    • Molecules and Cells
    • /
    • v.27 no.4
    • /
    • pp.391-396
    • /
    • 2009
  • Image-based, high-content screening assays demand solutions for image segmentation and cellular compartment encoding to track critical events - for example those reported by GFP fusions within mitosis, signalling pathways and protein translocations. To meet this need, a series of nuclear/cytoplasmic discriminating probes have been developed: DRAQ5$^{TM}$ and CyTRAK Orange$^{TM}$. These are spectrally compatible with GFP reporters offering new solutions in imaging and cytometry. At their most fundamental they provide a convenient fluorescent emission signature which is spectrally separated from the commonly used reporter proteins (e.g. eGFP, YFP, mRFP) and fluorescent tags such as Alexafluor 488, fluorescein and Cy2. Additionally, they do not excite in the UV and thus avoid the complications of compound UV-autofluorescence in drug discovery whilst limiting the impact of background sample autofluorescence. They provide a convenient means of stoichiometrically labelling cell nuclei in live cells without the aid of DMSO and can equally be used for fixed cells. Further developments have permitted the simultaneous and differential labelling of both nuclear and cytoplasmic compartments in live and fixed cells to clearly render the precise location of cell boundaries which may be beneficial for quantitative expression measurements, cell-cell interactions and most recently compound in vitro toxicology testing.

3D Quantitative Analysis of Cell Nuclei Based on Digital Image Cytometry (디지털 영상 세포 측정법에 기반한 세포핵의 3차원 정량적 분석)

  • Kim, Tae-Yun;Choi, Hyun-Ju;Choi, Heung-Kook
    • Journal of Korea Multimedia Society
    • /
    • v.10 no.7
    • /
    • pp.846-855
    • /
    • 2007
  • Significant feature extraction in cancer cell image analysis is an important process for grading cell carcinoma. In this study, we propose a method for 3D quantitative analysis of cell nuclei based upon digital image cytometry. First, we acquired volumetric renal cell carcinoma data for each grade using confocal laser scanning microscopy and segmented cell nuclei employing color features based upon a supervised teaming scheme. For 3D visualization, we used a contour-based method for surface rendering and a 3D texture mapping method for volume rendering. We then defined and extracted the 3D morphological features of cell nuclei. To evaluate what quantitative features of 3D analysis could contribute to diagnostic information, we analyzed the statistical significance of the extracted 3D features in each grade using an analysis of variance (ANOVA). Finally, we compared the 2D with the 3D features of cell nuclei and analyzed the correlations between them. We found statistically significant correlations between nuclear grade and 3D morphological features. The proposed method has potential for use as fundamental research in developing a new nuclear grading system for accurate diagnosis and prediction of prognosis.

  • PDF

An Image Cytometric MTT Assay as an Alternative Assessment Method of Nanoparticle Cytotoxicity

  • Lee, Song Hee;Park, Jonghoon;Kwon, Dongwook;Yoon, Tae Hyun
    • Bulletin of the Korean Chemical Society
    • /
    • v.35 no.7
    • /
    • pp.1933-1938
    • /
    • 2014
  • Despite increasing importance of in vitro cell-based assays for the assessment of nanoparticles (NPs) cytotoxicity, their suitability for the assessment of NPs toxicity is still in doubt. Here, limitations of widely used cell viability assay protocol (i.e., MTT asssay) for the cytotoxicity assessment of P25 $TiO_2$ NPs were carefully examined and an alternative toxicity assessment method to overcome these limitations was proposed, where the artifacts caused by extracellularly formed formazan and light scattered by agglomerated NPs were minimized by measuring only the intracellular formazan via image cytometric methods.

Utility of Nuclear Morphometry in Effusion Cytology

  • Ambroise, Marie Moses;Jothilingam, Prabhavati;Ramdas, Anita
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.16
    • /
    • pp.6919-6922
    • /
    • 2014
  • Background: The cytological analysis of serous effusions is a common investigation and yields important diagnostic information. However, the distinction of reactive mesothelial cells from malignant cells can sometimes be difficult for the cytopathologist. Hence cost-effective ancillary methods are essential to enhance the accuracy of cytological diagnosis. The aim of this study was to examine the utility of nuclear morphometry in differentiating reactive mesothelial cells from malignant cells in effusion smears. Materials and Methods: Sixty effusion smears consisting of 30 effusions cytologically classified as malignant (adenocarcinomas) and 30 benign effusions showing reactive mesothelial cells were included in the study. ImageJ was used to measure the nuclear area, perimeter, maximal feret diameter, minimal feret diameter and the circularity. A total of ten representative cells were studied in each case. Results: Significant differences were found between benign and malignant effusions for the nuclear area, perimeter, maximal feret diameter and minimal feret diameter. No significant difference was found for circularity, a shape descriptor. Receiver operating characteristic (ROC) curve analysis revealed that nuclear area, perimeter, maximal feret diameter, and minimal feret diameter are helpful in discriminating benign and malignant effusions. Conclusions: Computerised nuclear morphometry is a helpful ancillary technique to distinguish benign and malignant effusions. ImageJ is an excellent cost effective tool with potential diagnostic utility in effusion cytology.

Evaluation of Volumetric Texture Features for Computerized Cell Nuclei Grading

  • Kim, Tae-Yun;Choi, Hyun-Ju;Choi, Heung-Kook
    • Journal of Korea Multimedia Society
    • /
    • v.11 no.12
    • /
    • pp.1635-1648
    • /
    • 2008
  • The extraction of important features in cancer cell image analysis is a key process in grading renal cell carcinoma. In this study, we applied three-dimensional (3D) texture feature extraction methods to cell nuclei images and evaluated the validity of them for computerized cell nuclei grading. Individual images of 2,423 cell nuclei were extracted from 80 renal cell carcinomas (RCCs) using confocal laser scanning microscopy (CLSM). First, we applied the 3D texture mapping method to render the volume of entire tissue sections. Then, we determined the chromatin texture quantitatively by calculating 3D gray-level co-occurrence matrices (3D GLCM) and 3D run length matrices (3D GLRLM). Finally, to demonstrate the suitability of 3D texture features for grading, we performed a discriminant analysis. In addition, we conducted a principal component analysis to obtain optimized texture features. Automatic grading of cell nuclei using 3D texture features had an accuracy of 78.30%. Combining 3D textural and 3D morphological features improved the accuracy to 82.19%. As a comparative study, we also performed a stepwise feature selection. Using the 4 optimized features, we could obtain more improved accuracy of 84.32%. Three dimensional texture features have potential for use as fundamental elements in developing a new nuclear grading system with accurate diagnosis and predicting prognosis.

  • PDF