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At least 217 records · Page 12Linked to original sources
PubMed1986

Image cytometry in malignancy grading of breast cancer. Results in a prospective study with seven years of follow-up.

A fully automated system for image cytometry is presented. In combination with a special tumor sampling procedure, the system proved able to record with high reproducibility a large number of tumor cell population descriptors of high prognostic significance in a prospective study of breast cancer. These included the degree of abnormality of DNA distribution (strongly correlated with the proliferative rate) and some descriptors of shape and texture (strongly correlated with the grading system for breast cancer suggested by the World Health Organization). While the extreme complexity of a true malignancy grading system for breast cancer is underlined, the results of this study indicate that its creation is a possibility, given sufficient effort by interdisciplinary teams using modern sophisticated analytical techniques.

Breast Neoplasms↗
PubMed1998

Prognostic relevance of DNA image cytometry in oral cavity carcinomas.

OBJECTIVE: To evaluate the relation of DNA content of squamous cell carcinomas of the oral cavity to clinicopathologic features by image cytometry. STUDY DESIGN: A series of 52 patients with operable oral cavity carcinomas at stage T1-4 and NO-3 were studied. The tumors were classified according to the TNM classification and graded histopathologically. RESULTS: A positive correlation between tumor size and ploidy status was observed. There was a significant correlation between ploidy status and histologically proven existence of cervical lymph node metastases (61% nondiploid versus 29% diploid, P < .03). A relationship between histopathologic grade and ploidy status could not be proven. The cumulative five-year survival rate among patients with diploid DNA findings as opposed to those with nondiploid findings was significantly increased (P < .03). CONCLUSION: When assessing the prognosis of squamous cell carcinomas, DNA cytometry analysis can be used as an additional method. The significantly higher N stage, a higher frequency of metastases and a significantly lower survival rate with nondiploid tumors underline the importance of this examination method.

Adult↗
PubMed1998

Use of image cytometry to classify biliary and ampullary adenocarcinomas.

OBJECTIVE: To create an objective classification system to perform TNM classification of ampullary adenocarcinoma and cholangiocarcinoma using image cytometric data derived from Feulgen-stained tumor nuclei. STUDY DESIGN: Surgically resected cases of ampullary adenocarcinoma and cholangiocarcinoma with established TNM classifications were selected on the basis of available formalin-fixed, paraffin-embedded tissue. Fifteen numerical variables related to morphometric, densitometric and textural features of each tumor nucleus were recorded. We employed a methodology based on multivariate statistical tools to characterize the association of morphonuclear variables with TNM classification. The first step consisted of identifying and selecting representative nuclei of each T class. From this "purified" data set an objective classification system was created. The classification system was assessed using internal and external validation. RESULTS: Employing ANOVA, all 15 variables were significantly associated with T classification, 11 of 15 with N and 4 with M. Multivariate analysis was employed to distinguish between T1, T2 and T3 lesions. Our methodology correctly classified 76% of T1 nuclei, 47% of T2 nuclei and 84% of T3 nuclei. Heterogeneity within an individual tumor was defined in 61% of cases included in the training set. Complete concordance between pathologic classification and the classification system was observed in 71% of an independent validation.

Adenocarcinoma↗
PubMedDate not supplied

Image cytometry DNA analysis of diethylnitrosamine-induced dysplasias and invasive squamous cell carcinomas of the esophagus in mice.

The nuclear DNA content was assessed by image cytometry in squamous epithelial dysplasias and invasive squamous cell carcinomas of the esophagus induced by diethylnitrosamine (DEN) in mice. The study comprises 48 lesions: 27 lesions with low grade (i.e. slight and moderate) dysplasia, 18 with high grade dysplasia (i.e. severe dysplasia and CIS), and 3 with invasive squamous cell carcinoma. In addition, 5 parallel run control specimens were also investigated. The results demonstrated that only 3.7% (or 1/27) low grade dysplasias but as much as 72.2% (or 13/18) high grade dysplasias, and all three invasive squamous cell carcinomas displayed non-diploid DNA values. Three of 18 high grade dysplasias and all three invasive squamous cell carcinomas demonstrated aneuploid cell nuclei. The results of the present work indicate that the esophageal mucosa of the mouse permit investigation--under controlled conditions--of the nuclear DNA alterations occurring during carcinogenesis in this organ. The results presented herein thus substantiate the theory of increasing DNA aberrations occurring during carcinogenesis in the human esophagus.

Animals↗
PubMed1992

DNA ploidy and proliferation index of soft tissue sarcomas determined by image cytometry of fresh frozen tissue.

The DNA content of Feulgen-stained monolayer imprint preparations from 30 fresh frozen soft tissue and bone sarcomas was analyzed using image cytometry. DNA aneuploidy showed a significant association with increasing tumor grade. Of the grade III tumors, 83.3% (15 of 18) were aneuploid; of the grade I and II sarcomas, 33.3% (4 of 12) were aneuploid (P less than 0.00861). The proliferation index (PI) was determined by the percentage of tumor nuclear area staining with the monoclonal antibody Ki-67. The PI was significantly associated with ploidy and tumor grade. Ki-67 PI was elevated (greater than 2.0%) in 73.7% (14 of 19) of the aneuploid tumors, compared with 30% (3 of 10) of the euploid tumors (P less than 0.04597). Ki-67 PI was greater than 2.0% in 77.8% (14 of 18) of the grade III tumors, compared with 27.2% (3 of 11) of the grade I and II tumors (P less than 0.01773). These findings suggest that DNA ploidy and Ki-67 PI as determined by image analysis may be a useful supplement to tumor grading. The literature examining previous studies of sarcoma DNA ploidy is reviewed.

Adolescent↗
PubMed1987

A comparative study of Spitz nevus and nodular malignant melanoma using image analysis cytometry.

Image analysis cytometry can be used to estimate both nuclear DNA content and area in tissue sections. Since nodular malignant melanoma and Spitz nevus can show a remarkably similar light microscopic appearance, but may differ in behavior, we studied typical examples of these neoplasms to determine whether cytometric differences existed. Analysis of relative DNA content alone could not discriminate between these 2 entities in the 13 cases that we examined. However, Spitz nevi and nodular melanoma clearly differed in terms of maturation, which we defined as the difference between the mean nuclear size or mean nuclear DNA content of the uppermost and deepest melanocytes in each lesion. Maturation as defined by a decrease in mean nuclear DNA content proved highly significant (p less than 0.005). Only Spitz nevi showed a lesser DNA content in the deepest dermal cells as compared with upper dermal cells, suggesting that some Spitz nevi have an admixture of diploid and hyperdiploid cells in their upper portions, but mostly diploid cells in their deep portions. Only nodular melanoma showed higher mean DNA content in deep dermal cells as opposed to superficial dermal cells, suggesting that some nodular melanomas may either have clones of cells in their deep portions that have higher levels of ploidy, or more cells in the deep portion of melanomas may be in active phases of the cell cycle. Our study suggests that important cytometric differences exist between Spitz nevi and nodular melanoma, and that these could be exploited to develop cytometry into an adjunctive clinical technique.

Cell Nucleus↗
PubMed1992

Ploidy analysis of epithelial ovarian cancers using image cytometry.

We used a computerized image analysis system to determine the DNA content of 103 epithelial ovarian cancers using touch imprints of frozen tumor samples. Similar to prior studies of ploidy using flow cytometry, we found that most ovarian cancers (78%) were aneuploid while a minority (22%) were diploid. There was no relationship between ploidy and stage, histologic grade, or the ability to perform optimal cytoreductive surgery. Also, like prior studies using flow cytometry, negative second-look laparotomy and survival were somewhat more common in advanced-stage patients with diploid cancers than in those with aneuploid cancers. We conclude that ploidy of ovarian cancers can be determined using a computerized image analysis system to quantitate feulgen staining of cells in touch imprints. Ploidy is unlikely to play a role in treatment planning for patients with advanced-stage disease. Larger studies of patients with early-stage disease are needed, however, to determine whether ploidy is a more accurate means of predicting which patients are most likely to benefit from adjuvant therapy.

Aneuploidy↗
PubMed1994

A user's guide for avoiding errors in absorbance image cytometry: a review with original experimental observations.

The sources of errors which may occur when cytophotometric analysis is performed with video microscopy using a charged-coupled device (CCD) camera and image analysis are reviewed. The importance of these errors in practice has been tested, and ways of minimizing or avoiding them are described. Many of these sources of error are known from scanning and integrating cytophotometry; they include the use of white instead of monochromatic light, the distribution error, glare, diffraction, shading distortion, and inadequate depth of field. Sources of errors specifically linked with video microscopy or image analysis are highlighted as well; these errors include blooming, limited dynamic range of grey levels, non-linear responses of the camera, contrast transfer, photon noise, dark current, read-out noise, fixed scene noise and spatial calibration. Glare, contrast transfer, fixed scene noise, depth of field and spatial calibration seem to be the most serious sources of errors when measurements are not carried out correctly. We include a table summarizing all the errors discussed in this review and procedures for avoiding them. It can be concluded that if accurate calibration steps are performed and proper guidelines followed, image cytometry can be applied safely for quantifying amounts of chromophore per cell or per unit volume of tissue in sections, even when relatively simple and inexpensive instrumentation is being used.

Cytological Techniques↗
PubMed1997

Influence of debris and aggregates on image cytometry DNA measurement of archival tumors.

OBJECTIVE: To investigate, for seven samples, the influence of unwanted elements (e.g., remains of erythrocyte cell membranes, sliced nuclei, damaged nuclei and aggregates) on image cytometry DNA ploidy measurements. STUDY DESIGN: Two normal reference tissues (brain and breast), one breast cancer and four brain tumors were studied. For each sample, the influence of the different classes of debris on DNA ploidy histograms and indices was evaluated. RESULTS: The influence differs regarding each class of debris and the index to be evaluated. CONCLUSION: Strict and precise elimination of debris and aggregates is required. Moreover, strategies and efforts that must be applied to automated elimination of these unwanted elements must be a direct function of the bias they introduce into DNA ploidy measurements.

Aged↗
PubMed1991

Image cytometry of progesterone receptor expression during the cell cycle in the MCF-7 cell line.

Progesterone receptors (PR) appear to be distributed in a heterogeneous way in mammary tumor cells. The study presented here was designed to examine if heterogeneity of PR expression is cell-cycle dependent. Immunofluorescence techniques were used to label PR on the MCF-7 human breast cancer cell line and image cytometry was used to analyze the PR expression during G0 (Ki-67 antigen-negative cells), G1, S, and G2/M cell-cycle phases. A second PR, BrdU, and DNA analysis was performed to study PR expression in the S-phase (BrdU-positive cells). Our results show that PR synthesis occurs preferentially during the G0-G1 transition and that PR levels are constant during the G1-G2 transition. The PR expression appears to be cell-cycle related and may therefore explain the heterogeneity of PR expression. However, the possibility that PR heterogeneity may be linked to the existence of PR-negative subclones cannot be ruled out.

Breast Neoplasms↗
PubMed1993

Cell survival measurements at low doses using an automated image cytometry device.

The statistical precision of cell survival data measured at low doses can be greatly improved if the number of cells assayed at each dose point is known exactly. This paper describes an image cytometry technique that has been developed for this purpose. Treated cells are plated into tissue culture flasks, and after attachment their locations are determined with 98% accuracy by automated scanning procedures. These locations are revisited after an appropriate incubation period to assay for colony formation. Colonies may be scored either manually or using image data collected by the device. Additional information, such as heterogeneity of colony size, can also be obtained.

Animals↗
PubMed1997

Ki-67 labeling indices in non-small cell lung cancer: comparison between image cytometry and flow cytometry.

Expression of the proliferation-associated nuclear antigen Ki-67 was studied in 27 human non-small cell lung cancers (NSCLCs). We measured immunohistochemical Ki-67 labeling indices using image cytometry (ICM) and flow cytometry (FCM) and compared the indices determined with the two methods. Ki-67 labeling indices of tumor cells ranged from 2% to 59% with ICM, and from 3% to 56% with FCM, varying from case to case. There was a linear correlation of values between the two methods (r = 0.77, P < 0.05). To examine whether Ki-67 labeling indices represent tumor proliferative activity, we studied the relationship between the Ki-67 labeling index and the S-phase fraction determined by FCM. There was a positive correlation between the Ki-67 labeling index and the S-phase fraction (r = 0.91, P < 0.01). Comparison of Ki-67 labeling indices and clinicopathological parameters showed lower Ki-67 labeling indices in well-differentiated tumors than in moderate and poorly differentiated tumors (P < 0.05). No correlation was observed between Ki-67 labeling indices and p53 expression. It was concluded that the Ki-67 labeling index was in fact measured with both ICM and FCM, and that it represents tumor proliferative activity of NSCLCs.

Adenocarcinoma↗
PubMed1996

Persistent gestational trophoblastic disease: DNA image cytometry and interphase cytogenetics have limited predictive value.

DNA flow cytometry has shown a wider spectrum of DNA content in the complete hydatidiform mole (CM) than the originally reported diploidy. Conflicting results have been published about the relationship of DNA content and the occurrence of persistent gestational trophoblastic disease (PGTD). In the present study, 71 cases of CM and 4 cases of partial mole accompanied by PGTD and 100 cases of CM without PGTD were evaluated with DNA image cytometry for differences in DNA-ploidy pattern, expressed as the 2.5c and 5c exceeding rates. A pilot study of 20 cases of each group was performed using interphase cytogenetics to detect differences in the frequency of numerical chromosomal aberrations and in sex chromosome composition. For this purpose, DNA probes specific for the pericentromeric regions of chromosomes 1 and X and for the long arm of chromosome Y were incubated on 6-micron paraffin tissue sections. The results showed no differences between CMs with or without PGTD; DNA polyploidy occurred in 99% and 98% of cases, respectively; the 2.5c exceeding rate and 5c exceeding rate were 62.6 and 62.4, and 6.5 and 6.0, respectively. The frequency of numerical chromosomal aberrations as detected by interphase cytogenetics was 23.4 and 22.8%. An XY pattern was found in 3 of 20 cases of CM with PGTD and in 4 of 20 cases of CM without PGTD. The four cases of partial mole showed a DNA-ploidy pattern identical to that of a CM. For this reason, they would be better reclassified as CMs, despite the presence of nucleated red blood cells or amnion. Although nuclear atypia and corresponding increased DNA content is pronounced but variable in CMs, the occurrence of PGTD is not related to variations in quantitative DNA content nor to gross heterology or homology in sex chromosomes.

Adult↗
PubMed2007

DNA image cytometry is a useful adjunct tool in the prediction of disease outcome in patients with stage II and stage III colorectal cancer.

BACKGROUND: We assessed the prognostic value of the nuclear DNA content measured in the primary tumours of 123 patients with stage II or stage III colorectal cancer (CRC). METHODS: Isolated nuclei from paraffin sections were stained with the Feulgen reaction, and DNA was measured using a computer-assisted image analysis cytometry system. We applied 4 different approaches in the analysis of DNA histograms: the ABCDE approach, histogram range, peak evaluation and DNA cut-off values. RESULTS: Using the histogram range, the narrow range was rare (3.7%) in patients who died of disease (n = 28) as compared with 16.4% among those alive (n = 74; p = 0.017). Modal peak evaluation was a significant predictor of disease-free survival (DFS; Kaplan-Meier log-rank p = 0.0235). In the range evaluation, the 1st set (low-start gates) was a significant predictor of DFS (log-rank p = 0.0121), where disease recurrence was closely associated with the widest range (1.8->10c; c = haploid DNA content) gates. Recurrence-free survival was 3 times better in narrow-gate histograms than wide-range histograms (p < 0.03). The 1st set also proved to be a significant predictor of disease-specific survival (DSS; log-rank p = 0.0045), which was markedly better (77.8-90.0%) among the patients with the narrow-gate histograms. Grading of the histogram range into 2 categories (with 6.0c as cut-off), was a powerful predictor of both DSS (log-rank p = 0.0092) and 5-year DFS (p = 0.0106) in the whole series, and separately in stage III (but not stage II) disease, with p = 0.0131 and p = 0.0201, respectively. CONCLUSION: The DNA image cytometry with careful analysis of the histograms may provide valuable prognostic information in CRC, with potential clinical implications in patient management, particularly in predicting the patients at high risk for recurrence who should be considered as candidates for adjuvant therapy.

Aged↗
PubMedDate not supplied

The nuclear status of human sperm cells by TEM image cytometry: nuclear shape and chromatin texture in semen samples from fertile and infertile men.

Changes in nuclear size, shape, and chromatin texture during spermiogenesis and epididymal transport of human sperm were recently analyzed using transmission electron microscopy (TEM) image cytometry followed by multivariate statistical analysis of data. In the present study, this same methodology was used to investigate the nuclear morphology of spermatozoa in semen samples from fertile and infertile men. Analysis was carried out on a large series of micrographs of sections of sperm nuclei from a donor group with proven fertility and from a patient group with a mean infertility duration of 10 years with no obvious male or female infertility factors (only a slight decrease in the proportion of sperm heads with normal morphology was noted in routine semen tests). For the patient group, it was found that nuclei had a significantly less flattened shape (i.e., increased roundness as a consequence of increased thickness and decreased length). Furthermore, significant differences between donor and patient groups were found for most parameters of chromatin texture. In the patient group, chromatin was less condensed, and there was more homogeneous distribution of the different degrees of chromatin condensation. In addition, the organization of chromatin condensation and distribution along the major axis of the nucleus was found to be significantly different in the two groups. Stepwise linear discriminant analysis indicated a good classification rate of only 66% for nuclei of patients when using the eight major nuclear parameters, thus indicating the striking heterogeneity of nuclear morphology for both patient and donor groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Nucleus↗
PubMed1993

Comparative DNA image cytometry in imprint-, cytospin- and tissue section preparations of breast carcinoma.

Histologic sections, imprint preparations and cell preparations by enzymatic lysis from thick sections of cases of breast carcinomas were used to study total nuclear DNA distribution by image cytometry. Histologic sections are a prerequisite to unequivocally identify tumour cells as well as whole connective tissue cells used as a standard. While imprint preparations and lysis preparations are quite comparable with respect to distributional features, only one of them suffices to recognize these features, especially peaks at higher ploidies. Using this two-method approach to breast carcinomas, it was possible to distinguish a distribution separate from previously recognized distributions: a distinct triploid peak appears for predominantly high grade malignant ductal carcinomas.

Breast Neoplasms↗
PubMed1994

The value of DNA image cytometry for the cytological diagnosis of well-differentiated breast carcinomas and benign lesions.

Cytological evaluation of breast diseases can lead to false diagnosis if benign lesions show epithelial atypia or if carcinomas are orthoplasic and well differentiated. In order to determine the potential usefulness of image cytometry for discrimination between benign and malignant breast lesions, we studied 120 Feulgen-stained imprints, half from benign lesions with epithelial atypia and half from well-differentiated carcinomas and determined values of parameters expressing DNA content, morphometry and chromatin texture. Twenty-five out of the 60 carcinoma imprints were found to be DNA-aneuploid and could thus be easily distinguished from benign imprints, as, in our and other authors' experiences, aneuploidy very rarely occurs in benign breast lesions. When the remaining 35 carcinoma imprints were compared with 38 benign lesion imprints, randomly selected from the 60 cases studied, only four cases were misclassified after a linear discriminant analysis; the most discriminant parameters were those expressing the heterogeneity of DNA content and the chromatin texture. Thus, in cases where cytological diagnosis is difficult, image analysis of Feulgen-stained imprints may be a very useful tool for correctly discriminating benign from malignant cases.

Aneuploidy↗