Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Image Cytometry”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 541 records · Page 30Linked to original sources
PubMed1996

DNA ploidy and MYC DNA amplification in ovarian carcinomas. Correlation with p53 and bcl-2 expression, proliferative activity and prognosis.

There is increasing evidence that DNA ploidy is a prognostic factor in ovarian carcinomas, but it is uncertain whether MYC DNA amplification is an epiphenomenon of DNA nondiploidy or a distinct biological change with an impact on the clinical course of the disease. To clarify these issues we analysed DNA ploidy by flow and image cytometry and MYC copy number by polymerase chain reaction in archival material from ovarian carcinomas with known follow up. The results were compared with proliferative activity (Ki67 index) and p53 and bcl-2 expression. DNA cytometry revealed nondiploidy in 84 of 144 cases (58.3%). Nondiploidy was statistically significantly correlated with histological tumour type, histological grade, Ki67 index > 10%, FIGO stage, presence of residual tumour after debulking surgery and adverse postoperative outcome. Furthermore, DNA nondiploidy was associated with p53 accumulation. We found that 84.9% of the p53-positive cases were nondiploid. This points to the paramount importance of wild type p53 for the maintenance of genome integrity in this tumour type. MYC DNA amplification was seen in 33.8% (26/77 cases) of ovarian carcinoma. There was no correlation between MYC DNA amplification and histological tumour type, histological grade, FIGO stage, DNA ploidy, proliferative activity or prognosis. However, when p53 and bcl-2 expression was taken into account, a statistically significant correlation between gene alteration or expression patterns and histological tumour type was revealed. The group of mucinous carcinomas demonstrated both MYC DNA amplification and strong bcl-2 expression in 50% and contained the largest fraction of cases without aberration (37.5%). Endometrioid carcinomas were characterized by strong bcl-2 expression in 85%, whereas serous and undifferentiated carcinomas predominantly exhibited p53 alterations, frequently accompanied by bcl-2 overexpression or MYC DNA amplification. Thus, in interaction with other genes MYC DNA amplification may play a role in the determination of the varying differentiation patterns of ovarian carcinomas.

Aged↗
PubMed1994

Comparison of image (CAS 200) and flow cytometry determined DNA content of paraffin-embedded Hodgkin's disease tissue.

To assess the reliability of DNA estimation using image cytometry, deparaffinized lymph nodes from 70 patients with Hodgkin's disease were examined and the results obtained were compared with those from flow cytometry. Image analysis without discriminating between the various cell types, as found in Hodgkin's disease, revealed no separate aneuploid peak. Selecting on morphologically defined nuclear types DNA aneuploidy was detected in 20% of the cases (14/70). The aneuploid populations were limited to the population of nuclei defined as Reed-Sternberg (RS)-like or medium-sized lymphocytes. Benign lymph nodes DNA aneuploidy was not found in any of the controls. Comparison of DNA histograms obtained by image and flow cytometry showed aneuploid peaks using image cytometry in 4 of 30 diploid and 10 of 40 aneuploid flow histograms. In conclusion, image analysis using the CAS 200 system as compared to flow cytometry is more time-consuming and less sensitive to assess ploidy status, although it may provide extra information in some selected cases. Evidence is obtained that DNA aneuploidy in Hodgkin's disease is preferentially expressed by cells with the RS/H-like and medium-sized lymphocyte morphology.

Aneuploidy↗
PubMed1997

Laser scanning cytometry distinguishes lymphocytes, monocytes, and granulocytes by differences in their chromatin structure.

By providing rapid measurements of cellular fluorescence in addition to morphometric analysis, the novel microscope-based multiparameter laser scanning cytometer (LSC) combines advantages of flow and image cytometry. Analysis of the integrated fluorescence intensity (IF) versus peak fluorescence intensity (maximal fluorescence per pixel; FP) versus fluorescence area (FA) of the cells stained with the DNA intercalating fluorochrome propidium iodide (PI) made it possible to discriminate lymphocytes, monocytes, and granulocytes in samples of peripheral blood of normal individuals. Lymphocytes, characterized by maximally condensed chromatin, had the highest FP and lowest values of FA. Granulocytes had the lowest FP and the highest FA. They also had increased IF compared to lymphocytes and monocytes. The difference in IF between granulocytes and monocytes/lymphocytes was abolished after exposure of cells to 0.1 M HCl at 0-4 degrees C, which is known to dissociate histones from DNA in chromatin. Monocytes were characterized by intermediate values of peak and area fluorescence intensity compared to lymphocytes and granulocytes. Thus, although all three classes of white blood cells have the same DNA content, they can be distinguished based on differences in structure of their chromatin after staining with PI. Discrimination of these cells by LSC is similar to that provided by flow cytometry based on differences in forward and side light scatter properties.

Cell Separation↗
PubMed1998

Malignancy associated changes in bronchial epithelial cells and clinical application as a biomarker.

A total of 74 bronchial brushing specimens, 24 from patients with advanced stage cancer, eight from patients with CIS, 31 from patients with atypical metaplasia and 11 from normal subjects were examined for the existence of malignancy associated changes (MAC). Conventional fiberoptic bronchoscopy and fluorescence endoscopy was carried out on every case. Each case was classified according to the highest grade of abnormality diagnosed by bronchial biopsy of the suspect areas. During the endoscopy examination, a bronchial brushing specimen was obtained from a visually normal area very remote from the abnormal area as possible such as the opposite lung or another lobe. The bronchial brushing specimens were fixed, mounted and stained by a DNA specific method and approximately 1500 images of individual nuclei per case were captured by an automated high resolution image cytometry. For each of these images, more than 100 nuclear features such as size, shape and chromatin spatial organization were calculated. Discriminant function analysis revealed nuclear features which differentiated between normal bronchial cell nuclei from the normal subjects and ostensively normal nuclei (MAC cell nuclei) from the lung cancer patients. The best discrimination was achieved when the frequency of individual cells expressing MAC was 50% or greater. With this threshold, 75% of the patients with invasive cancer and CIS were correctly classified. Fifty percent of those with severe or moderate atypia and 35% with mild atypia were also MAC positive. The frequency of cells expressing MAC also increased as the degree of abnormality of the groups increased. MAC may be a useful criterium to determine biological behavior of the intra-epithelial (pre-invasive) neoplasia.

Aged↗
PubMed2001

Fetal adenomas and minimally invasive follicular carcinomas of the thyroid frequently display a triploid or near triploid DNA pattern.

The ploidy pattern and the percentage of S-phase cells were investigated by means of flow cytometry using fresh or frozen samples in a series of 143 tumors and tumor-like lesions of the thyroid in an attempt to find whether there is any relationship between the histological characteristics of the lesions and their DNA content. The percentages of aneuploidy cases per category were: nodular goiter, 18.5% (15/81); fetal adenoma (including cases with trabecular/solid growth pattern), 58.3% (14/24); follicular adenoma other than fetal adenoma, 0% (0/18); papillary carcinoma, 11.1% (1/9); and minimally invasive follicular carcinoma, 57.1% (4/7). Regardless of the histological category, aneuploid lesions had a significantly higher (P < 0.001) percentage of S-phase cells (7.3%) than diploid lesions (4.1%). All the six cases with a DNA content within the triploid range were fetal adenomas, but one was a follicular carcinoma displaying a fetal adenoma-like growth pattern. The other three follicular carcinomas with an aneuploid DNA pattern also displayed foci of fetal adenoma-like growth pattern. Image cytometry of the four aneuploid follicular carcinomas showed similar DNA indexes in the peripheral, invasive foci of the lesions and in the central fetal adenoma-like areas. These results demonstrate that aneuploidy in benign tumors is restricted to adenomas displaying a fetal or fetal/embryonal growth pattern and support the concept that chromosome instability is a major pathway of tumorigenesis in thyroid follicular neoplasms.

Adenocarcinoma, Follicular↗
PubMed1987

Development of an imaging flow cytometer.

A cell analyzer that combines the characteristics of image cytometry and flow cytometry is being designed and constructed at the University of Sydney. This paper describes the image acquisition and processing components and some preliminary applications. Cells stained by a fluorescent dye and suspended in a liquid medium are conveyed by a hydraulic system to a flow channel assembly, where they are detected and illuminated by a laser beam. A two-dimensional charge-coupled device is used to acquire the cell images. Image processing and classification is to be carried out by a special-purpose computer comprising an array of four conventional microprocessors and a highly parallel processor consisting of an array of 32 X 32 processing elements. The analyzer will be capable of using morphologic, immunologic and biochemical information to classify and sort up to 500 cells per second. Because of its unique characteristics, the instrument will be of particular use in tumor heterogeneity studies.

Flow Cytometry↗
PubMed1994

DNA ploidy measurements in prostate cancer: differences between image analysis and flow cytometry and clinical implications.

The relationship between DNA ploidy and prognostic parameters has been established in prostate cancer. However, comparison of various techniques used for DNA ploidy analysis has not been sufficiently addressed in prostate carcinoma. In the present study, grossly identified discrete carcinoma foci from 48 consecutive radical prostatectomy specimens were analyzed by flow cytometry and image analysis using both imprints and tissue sections. Correlations with other prognostically important pathological findings such as grade, size, extracapsular extension, positive surgical margin, and seminal vesicle and lymph node involvement were done. Image analysis detected 26 (54.2%) nondiploid tumors compared to 15 (31.3%) nondiploid tumors by flow cytometry. No significant differences were demonstrated between imprints and tissue sections. DNA ploidy status showed a good correlation with most other pathological findings listed above. In conclusion, DNA ploidy status determination gives clinically useful information which correlates well with the other useful pathological parameters of prognostic importance. Image analysis may be more sensitive than flow cytometry in detecting nondiploid populations. Tissue sections appear to be as reliable as imprints when properly controlled.

Adenocarcinoma↗
PubMed1992

Time dimension in histopathology.

In the liver acinus hepatocytes and littoral cells stream from the portal tract toward the terminal hepatic vein. Their average displacement trajectory was denominated as tissue radius. Since cells advance on the tissue radius in one direction, the farther a cell is the older it is, and cell age may be estimated from its position. This property is regarded as time dimension of a tissue. Since distance may be estimated by image cytometry, age determination may be incorporated in the image cytometer software. Distance may be expressed in two types of units: 1. metric e.g. microns, and 2. cell location or the number of cells separating a given cell from tissue origin. Cell age may also be expressed by two units: chronological and biological. Chronological age may be derived from the cell's displacement velocity. A hepatocyte advances daily 2 microns so that a cell at 100 microns distance is 50 days old. Biological age is defined as the cell's location on the radius. Cell location may serve as biological age unit since cells ranked according to their location are also ranked by their ages. The average acinus radius is 22 location long so that in biological age units, maximal hepatocyte life span is 22 locations. One may even say that a cell at location 5 is 2 biological units older than a cell at location 1. In order to estimate chronological age the cell has to be followed for a period of time, while biological age may be read off once from the section. There is yet another advantage for utilizing cell location as unit.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Division↗
PubMed2001

Cyclin A and Ki-67 with DNA content in benign and malignant prostatic epithelial lesions.

OBJECTIVE: To evaluate the usefulness of immunohistochemical staining of cyclin A and Ki-67 together with DNA content in the classification of benign prostatic hyperplasia, prostatic intraepithelial neoplasm (PIN) and prostatic carcinoma foci and to compare these parameters with each other and with parameters obtained from conventional histopathology. STUDY DESIGN: We selected 37 carcinoma, 18 PIN and 8 hyperplastic foci from prostatectomies done during 1996 and 1997 at Turku University Central Hospital. Cyclin A and Ki-67 staining was assessed by immunohistochemistry and DNA content by image cytometry. RESULTS: The hyperplastic, PIN and carcinoma foci differed clearly in their 2.5c exceeding rates, image cytometric proliferation indices and staining indices for cyclin A and Ki-67. No significant differences were found between these histologic entities in their modal DNA ploidy values. In carcinomas, cyclin A and Ki-67 indices differed between low, intermediate and high Gleason and World Health Organization grading groups. Diploid and tetraploid carcinomas had similar cyclin A and Ki-67 indices, which differed from those of aneuploid carcinomas. CONCLUSION: The 2.5c exceeding rate and image cytometric proliferation index as well as the cyclin A and Ki-67 indices differed significantly between different types of prostatic lesions. Cyclin A and Ki-67 had good correlations with the histologic grade of carcinoma.

Biomarkers, Tumor↗
PubMed1988

Application of flow cytometry and automated image analysis to the study of prostate cancer.

Flow cytometry and image analysis are complementary quantitative cytologic techniques that have demonstrated utility in the assessment and analysis of prostate cancer and other urologic and nonurologic tumors. This review is intended to assess the current state of the art and to project future directions and applications of these modalities in the pathologic assessment of prostate cancer. Special attention is directed toward the topics of prostate cancer detection and diagnosis, determination of patient prognosis, and the monitoring of patient response to therapy. We recommend that 1) flow cytometry and image analysis be used to determine pathologic parameters that will help in predicting poor patient prognosis and 2) quantitative cytologic determinations of DNA content be included in clinical trials so that their ultimate role in monitoring patient response to therapy can be determined. This knowledge will allow the development of protocols designed to test the value of earlier institution of multimodal therapy in high-risk populations.

Flow Cytometry↗
PubMed1996

Retrospective DNA ploidy analysis by image and flow cytometry in head and neck cancer.

A retrospective analysis of DNA content (DNA ploidy) was conducted in formalin-fixed specimens from patients with advanced, unresectable head and neck cancer who were treated on a single defined protocol with accelerated fractionation radiation therapy and concomitant cisplatin (Platinol) chemotherapy. Specimens from 31 tumor sites were analyzed by image cytometry using the Feulgen staining method. Fifteen specimens were analyzed by flow cytometry after deparaffinization, nuclear disaggregation, and staining with propidium iodide. By image analysis, 10 (32%) of 31 specimens contained only diploid tumor cells, while 21 (68%) specimens exhibited at least one aneuploid tumor component. Seven of the eight tumors with a single G0/G1 peak by image analysis had a single peak by flow analysis. Five of the seven tumors with multiple peaks by image analysis had multiple peaks by flow analysis. Histology was also reevaluated, and tumor grade was determined, reflecting tumor cell differentiation based on keratinization, mitotic activity and the degree of nuclear polymorphism. For this well-defined patient population managed according to a uniform therapeutic approach, DNA ploidy status and histology provided a suggestion of prognostic separation; however, statistical significance was not obtained, most likely due to the small number of patients.

Combined Modality Therapy↗
PubMed2003

Applicability of liquid-based cytology to the assessment of DNA content in cervical lesions using static cytometry.

OBJECTIVE: To evaluate the nuclear DNA content of cervical lesions in liquid-based cytologic specimens prepared for static cytometry. STUDY DESIGN: The DNA content of cervical lesions was evaluated in cervical samples prepared with the Autocyte PREP liquid-based cytology system (TriPath Imaging Inc., Burlington, North Carolina, U.S.A.). A series of 47 samples stained with the Papanicolaou method (chronic cervicitis, n = 15; cervical intraepithelial neoplasia [CIN] 1, n = 25; CIN 2, n = 5; CIN 3, n = 2) were collected from consecutive women enrolled in an ongoing screening study at Leonor Mendes de Barros Hospital, São Paulo, Brazil, in 2002. Each residual sample was processed according to the Feulgen-thionin method (TriPath Imaging). Ploidy evaluation was performed using the CAS 200 image analysis system and Quantitative DNA Measurement software 3.0 (version 8.1) (Becton Dickinson, San Jose, Califoria, U.S.A.). Cellular ploidy was analyzed from atypical nuclei, and the DNA index was obtained using histograms for interpretation. RESULTS: All chronic cervicitis cases were diploid. Of the CIN 1 cases, 44% were diploid, 12% tetraploid, 32% aneuploid and 12% polyploid (diploid plus tetraploid). CIN 2 lesions were diploid in 60% and aneuploid in 40% of cases, whereas all CIN 3 lesions (100%) were aneuploid. CONCLUSION: The liquid-based cytologic samples proved to be suitable and highly useful for DNA analysis by image cytometry, which was capable of discriminating CIN 3 lesions from CIN 1 and 2 but not CIN 1 from 2 lesions. Aneuploidy was closely associated with CIN 3 lesions.

DNA↗
PubMed1998

DNA-cytometry of progressive and regressive cervical intraepithelial neoplasia.

A retrospective analysis was performed on archival cervical smears from a group of 56 women with cervical intraepithelial neoplasia (CIN), who had received follow-up by cytology only. Automated image cytometry of Feulgen-stained DNA was used to determine the differences between progressive and regressive lesions. The first group of 30 smears was from women who had developed cancer after initial smears with dysplastic changes (progressive group). The second group of 26 smears with dysplastic changes had shown regression to normal (regressive group). The goal of the study was to determine if differences in cytometric features existed between the progressive and regressive groups. CIN categories I, II and III were represented in both groups, and measurements were pooled across diagnostic categories. Images of up to 700 intermediate cells were obtained from each slide, and cells were scanned exhaustively for the detection of diagnostic cells. Discriminant function analysis was performed for both intermediate and diagnostic cells. The most significant differences between the groups were found for diagnostic cells, with a cell classification accuracy of 82%. Intermediate cells could be classified with 60% accuracy. Cytometric features which afforded the best discrimination were characteristic of the chromatin organization in diagnostic cells (nuclear texture). Slide classification was performed by thresholding the number of cells which exhibited progression associated changes (PAC) in chromatin configuration, with an accuracy of 93 and 73% for diagnostic and intermediate cells, respectively. These results indicate that regardless of the extent of nuclear atypia as reflected in the CIN category, features of chromatin organization can potentially be used to predict the malignant or progressive potential of CIN lesions.

Adult↗
PubMed1999

DNA-ploidy analysis correlates with the histogenetic classification of thymic epithelial tumours.

The ploidy values of the epithelial component were determined in a series of thymomas and organotypic thymic carcinomas using image cytometry and the results were compared with the histological tumour subtypes according to the histogenetic classification introduced by Marino, Müller Hermelink, and Kirchner (MMHK). Forty-six cases of thymic epithelial tumours were included in the study. After reclassification according to the MMHK classification, the distribution among the subtypes was as follows: three medullary, nine mixed type, five predominantly cortical (organoid), 16 cortical thymomas, and 13 well-differentiated thymic carcinomas. Single cell preparations were made from paraffin-embedded tumour tissue and stained according to Feulgen. Ploidy analysis was performed using an automated image analysis system. In five cases, DNA cytometry could not be performed, for technical reasons. The remaining 41 cases consisted of 11 diploid and 30 non-diploid tumours. The percentage of aneuploid tumours in the different subtypes increased from medullary (0 per cent) through mixed type (44.4 per cent), predominantly cortical (75 per cent), cortical (83.3 per cent) to well-differentiated thymic carcinomas (100 per cent). DNA-ploidy determination using image cytometry correlates with the concept of the MMHK classification of thymomas.

Humans↗
PubMed1998

Biclonal primary plasma cell leukemia.

Authors present a multiparameter pathological study in a case of rapid biclonal primary plasma cell leukemia. The immunohistochemical data revealed aberrant phenotypes (monocyte, epithelial and T-cell) probably in connection with microenvironmental influences. Biclonality can be attributed to class switching during malignant transformation. Static image cytometry showed aneuploidy. The blasts of this process are active immunoregulatory cells.

Adult↗
PubMed1997

Prognostic value from DNA quantification by static cytometry in breast cancer.

OBJECTIVE: To determine if DNA quantification on fine needle aspiration (FNA) has predictive value in breast cancer. STUDY DESIGN: Forty-nine patients with breast cancer were selected for this study because of their common characteristics. The smears were studied using image cytometry. RESULTS: With the type of histogram and value of entropy, two large groups, with high and low degrees of malignancy, were obtained. Survival was utilized as a variable of interest. Kaplan-Meier survival curves for both groups were formulated, and the results were supported with statistical data. CONCLUSION: The survival differences between both groups were statistically significant (P < .001), thus demonstrating the predictive value of DNA quantification.

Biopsy, Needle↗
PubMed2001

Nasopharyngeal carcinoma heterogeneity of DNA content identified on cytologic preparations.

OBJECTIVE: To evaluate tumor heterogeneity of DNA content in nasopharyngeal carcinoma (NPC) performed on cytologic specimens. STUDY DESIGN: Image cytometric analysis of DNA ploidy status of 40 NPCs was performed on nasopharyngeal brushing smears stained with the Feulgen method after hematoxylin eosin staining. If the DNA distribution pattern from the same tumor exhibited diploid, aneuploid or/and tetraploid peaks or some combination of these patterns, the presence of tumor heterogeneity of DNA content was identified. RESULTS: Thirty-four cases (85%) had a nondiploid DNA pattern among the 40 NPCs. Twenty-eight cases exhibited tumor heterogeneity of DNA content (70%). Of the 28 tumors, 13 (46%) had a combination of diploid and tetraploid patterns, 10 (37%) had a combination of diploid and aneuploid patterns, 3 cases (11%) had a combination of tetraploid and aneuploid patterns, and 2 cases had two aneuploid stem lines. The relationship between DNA ploidy pattern and tumor histologic and cytologic morphology was also examined. CONCLUSION: There is a high incidence of DNA content heterogeneity in NPC. The relevance of tumor heterogeneity to the biologic behavior of NPC awaits further study. DNA quantification with image cytometry on destained cytologic preparations is feasible and reliable.

Aneuploidy↗
PubMed1990

Nuclear DNA distribution pattern of the parenchymal cells in adenocarcinomas of the pancreas and in chronic pancreatitis. A study of archival specimens using both image and flow cytometry.

The nuclear DNA distribution pattern of the neoplastic parenchymal cells of 100 conventionally formalin-fixed and paraffin-embedded specimens from pancreatic adenocarcinomas and from 8 specimens of chronic pancreatitis was assessed by means of image cytometry. All material originated from pancreatic restrictions. Evaluable DNA histograms could be obtained for 77 carcinomas, and clinical data were available for 71 of these. In these 71 specimens, the nuclear DNA ploidy pattern was also investigated by means of flow cytometry. In 76 of the 77 cases, the image-cytometric DNA ploidy pattern obtained showed a "nondiploid" distribution with modal values as high as 8.5 c. In 21 cases, the neoplastic cells showed modal values in the "triploid" region. The analogous 71 flow-cytometric DNA histograms could only be evaluated in 50 cases because of excessively high amounts of background and/or excessively broad peaks. In 47 cases, the nuclear DNA histogram was nondiploid according to both techniques. The patients with carcinomas whose cell nuclei showed a triploid DNA distribution showed a significantly shorter survival time than those with tumor cell populations of nontriploid DNA distribution patterns. In the 8 specimens of chronic pancreatitis, the parenchymal cells were all equipped with nuclei showing diploid DNA distribution patterns.

Adenocarcinoma↗