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DNA aneuploidy as a specific marker of neoplastic cells in FNAB of the thyroid.

OBJECTIVE: To investigate whether DNA image cytometry (ICM) could be of value in the specific identification of neoplastic cells in cytologic specimens of the thyroid. STUDY DESIGN: FNABs of thyroid from 162 patients with different benign and neoplastic diseases were investigated. Nuclear DNA content in thyroid cells was measured after Feulgen staining using a TV image analysis system. Data were correlated with clinical and histologic patient follow-up. The occurrence of abnormal DNA stemlines was used as a marker for aneuploidy and thus for neoplasia. RESULTS: None of the 89 cases without tumor cells revealed DNA aneuploidy. An abnormal DNA stemline was found in 55% of histologically proven benign thyroid tumors (follicular or oncocytic adenomas) and in 59.5% of malignant tumors. The positive predictive value of DNA aneuploidy in FNABs of the thyroid for neoplasms was 100%. The negative predictive value of DNA nonaneuploidy for the prediction of tumor-free histologic or clinical follow-up was 79.4%. CONCLUSION: DNA image cytometry may be helpful in the specific identification of neoplastic follicular cells. DNA ICM on FNABs of the thyroid is an additional tool to achieve early identification of patients with nodular lesions of the thyroid that have to be operated on. DNA euploidy excludes the presence of neither malignancy nor neoplasia.

Adolescent↗

Image cytometric DNA analysis of adrenocortical neoplasms as a prognostic parameter: a clinico-pathologic study of 13 patients.

Formalin-fixed paraffin-embedded material of thirteen adrenocortical neoplasms were examined by conventional microscopy, and by DNA image cytometry in Feulgen-stained tissue sections and cytospin preparations. The subsequent clinical course of each patient was reviewed. Seven of the neoplasms were classified according to the histologic criteria of Weiss as adenomas and six as carcinomas. The DNA content and indices of all 13 adrenocortical neoplasms were measured by image cytometry. For the internal standard, the DNA values of the adjacent normal adrenal cells in 10 cases and of nuclei of fibrocytes of the tumour stroma were determined. DNA values in all seven adenomas, in normal adrenal cells, and in the nuclei of fibrocytes were unequivocally diploid. The six carcinomas expressed aneuploid, tetraploid or hypertetraploid DNA values. DNA values of normal adrenal cells seemed to provide a better internal control than those of connective tissue cells. Of the seven adenoma patients with a follow-up between 8 and 124 months (mean 59 months), six were alive without evidence of tumour; one patient died from other causes. Of the six carcinoma patients with a follow-up ranging from 5 days to 36 months, three patients died from the disease; one patient was alive with metastases, and only two patients were alive without evidence of the disease for 25 and 36 months, respectively. Our results strongly suggest that image cytometric DNA analysis is a valuable method to distinguish between benign and malignant adrenocortical neoplasms.

Adrenal Cortex Neoplasms↗

Nuclear chromatin texture and sensitivity to DNase I in human leukaemic CEM cells incubated with nanomolar okadaic acid.

It is now known that the analysis of chromatin texture can be used in oncology as a sensitive detection method, either to define diagnostic classifications or to locate a lesion along a defined trend curve. However, the functional significance of these variations in textural features remains sometimes unclear. Several drugs have been shown to be able to modulate chromatin structure. Among them, the phosphatase inhibitor okadaic acid at low concentration can increase accessibility to DNA in chromatin of carcinoma cells. This paper demonstrates that short exposures (0-3 h) to a 10-nM dose of okadaic acid induced an increased sensitivity to DNase I digestion in human CEM leukaemic cell nuclei and that this sensitization was associated to variations of nuclear texture characteristics, as evaluated by image cytometry. CEM cells treated with okadaic acid for 0-3h displayed changes in chromatin supraorganization with a more homogeneous and fine chromatin texture, as compared to control cells. This suggests that the appearance of an open configuration of chromatin structure as evaluated by biochemical methods corresponds to a more decondensed texture of nuclei measured by image cytometry. Longer exposures (6-24h) of CEM cells to 10 nM okadaic acid lead to apoptosis. As reported previously for camptothecin-treated HL60 cells, okadaic acid-treated CEM cells display biphasic nuclear chromatin texture changes, i.e. a decondensation phase followed by the appearance of typical apoptotic cells with a smaller nuclear area and a highly condensed chromatin. Finally, using the multidrug-resistant CEM-VLB cell line, it was confirmed that these multidrug-resistant cells also display cross-resistance to okadaic acid, as this compound was unable to induce either increased DNase I sensitivity, apoptosis, or altered nuclear texture in this particular cell line.

Apoptosis↗

DNA quantification as a prognostic factor in colorectal adenocarcinoma.

OBJECTIVE: To determine if DNA quantification, studied in cytologic samples obtained by fiberendoscopy, has predictive value in colorectal adenocarcinoma. STUDY DESIGN: Thirty patients with colorectal cancer diagnosed by cytology, endoscopy and microbiopsy, were selected. The study covered > 10 years. Smears were stained with progressive hematoxylin and processed by computer for DNA evaluation by image cytometry. RESULTS: Four different types of histograms that directly related to malignancy were obtained. These histograms were characterized by the value of entropy, and we established four grades of aggressiveness. Finally, we obtained two large groups: high and low grade of malignancy. We studied the survival times in both groups and used a Kaplan-Meier survival curve for the high grade group. We confirmed the results statistically and found that there was a significant relationship, with P < .05. CONCLUSION: We strongly advise the use of DNA quantification by image cytometry in daily surgery as a prognostic indicator of survival time in colorectal cancer.

Adenocarcinoma↗

Impact of the preparation technique on DNA content measured by image analysis in early stage human testis cancer.

Current clinical staging which includes serum tumour markers and imaging techniques fails to identify 30-40% of clinical stage I nonseminomatous germ cell testicular tumour (NSGCT) patients who have occult metastatic disease at time of orchiectomy and who will, therefore, develop clinically evident metastases, usually within two years of follow-up. Therefore, there is a real clinical need to evaluate new biological parameters of the primary tumour which might be useful as predictors for occult metastatic disease. Some investigators have described that DNA content measured by image cytometry is of prognostic value in early stage NSGCT to detect patients at risk for occult metastatic disease. However, optimal preparatory techniques are mandatory in establishing new tumour biological markers in order to obtain reliable and reproducible results. This study has analyzed the impact of the sedimentation technique in comparison to the cytocentrifugation technique on DNA measurement in early stage NSGCT obtained by image cytometry. Different tissue blocks of formalin fixed, paraffin embedded primary testicular tumours (NSGCT) of 50 clinical stage I patients, who underwent retroperitoneal lymph node dissection between 1985 and 1989, were analyzed. Thirty (60%) patients had histologically proven lymph node involvement (pathological stage B), whereas 20 (40%) patients (pathological stage A) had neither lymph node metastases nor tumour recurrence during follow-up. The samples were prepared according to a modified Hedley technique: Individual tissue digestion times were monitored closely to avoid overdigestion. The times varied from 30 to 60 min depending on the constituents of the tissue section. Prolonged digestion times did not correlate with poor quality of the preparations and brief digestion times did not always yield optimal specimens. The impact of two different techniques (cytocentrifugation and gravity sedimentation) on the Feulgen staining results were compared. Cytocentrifuged samples consistently provided larger and paler nuclei with less well defined borders compared to slides from the same cell suspension processed by the sedimentation technique. Nuclei from pathologic stage II samples were more vulnerable to cytocentrifuge alteration than those of stage I. According to the results obtained in this study, the sedimentation slide preparation technique should be preferred for DNA ICM in NSGCT, and possibly in other human malignancies as well.

DNA, Neoplasm↗

Near-tetraploidy in adult acute myelogenous leukemia.

Tetraploidy and near-tetraploidy are observed infrequently in hematologic malignancies, most commonly seen in cases of childhood acute lymphoblastic leukemia, and are associated with large blast size. Four cases of adult acute myelogenous leukemia (AML) with tetraploid or near-tetra-ploid karyotypes are reported, along with review of the related literature. AML subtypes included M1, M1, M4, and M5b. Tetraploidy was determined cytogenetically and confirmed by image cytometry (DNA index 2.0). The subjective impression of large blast size was confirmed by image cytometry, demonstrating mean blast nuclear areas of 237, 177, 203, and 216 microns2, (mean 208 microns2) in the cases with tetraploidy, compared to a mean of 134 microns2 in 10 control cases of AML with diploid or near diploid chromosome patterns. The clinical course was variable in the four cases reported. When compared with previously published cases, the occurrence of tetraploidy or near-tetraploidy in adult AML, unlike childhood ALL, does not appear to define a distinct subgroup in terms of FAB classification or to carry prognostic implications.

Adult↗

Cytologic and DNA-cytometric early diagnosis of oral cancer.

OBJECTIVE: The aim of this prospective study was to report on the diagnostic accuracy of conventional oral exfoliative cytology taken from white-spotted, ulcerated or other suspicious oral lesions in our clinic. In addition we checked DNA-image cytometry as an adjuvant diagnostic tool. Our hypothesis is that DNA-aneuploidy is a sensitive and specific marker for the early identification of tumor cells in oral brushings. STUDY DESIGN: 251 cytological diagnoses obtained from exfoliative smears of 181 patients from macroscopically suspicious lesions of the oral mucosa and from clinically seemingly benign oral lesions which were excised for establishing histological diagnoses were compared with histological and/or clinical follow-ups of the respective patients. Additionally nuclear DNA-contents were measured after Feulgen restaining using a TV image analysis system. RESULTS: Sensitivity of our cytological diagnosis on oral smears for the detection of cancer cells was 94.6%, specificity 99.5%, positive predictive value 98.1% and negative predictive value 98.5%. DNA-aneuploidy was assumed if abnormal DNA-stemlines or cells with DNA-content greater 9c were observed. On this basis the prevalence of DNA-aneuploidy in smears of oral squamous cell carcinomas in situ or invasive carcinomas was 96.4%. Sensitivity of DNA-aneuploidy in oral smears for the detection of cancer cells was 96.4%, specificity 100%, positive predictive value 100% and negative 99.0%. The combination of both techniques increased the sensitivity to 98.2%, specificity to 100%, positive predictive value to 100% and negative to 99.5%. CONCLUSIONS: Brush cytology of all visible oral lesions, if they are clinically considered as suspicious for cancer, are an easily practicable, cheap, non-invasive, painless, safe and accurate screening method for detection of oral precancerous lesions, carcinoma in situ or invasive squamous cell carcinoma in all stages. We conclude that DNA-image cytometry is a very sensitive, highly specific and objective adjuvant tool for the early identification of neoplastic epithelial cells in oral smears.

Adult↗

[DNA quantitative analysis of head and neck tumor].

OBJECTIVE: This research discussed the diagnostic value of cellular quantitative analysis in head and neck tumor. METHOD: We analysis quantitative parameters of 23 malignant tumors and 20 benign diseases with automated DNA image cytometry, they are the DNA Index (DI), S-phase Fraction (SPF), Proliferation Index (PI) and 5c Exceeding (5c Ex). RESULT: The masculine rate of cellular quantitative analysis and conventional method are 76.92% and 50.00%. The value of SPF, PI and 5c Ex are 14.05%, 19.59% and 1.59% in malignant tumor, respectively. The parameters of SPF and PI showed distinct difference in the area of malignant tumor than the 1 cm sideward area of tumor and the edge of tumor. CONCLUSION: Automatic DNA image cytometry has higher masculine rate of malignant tumor than conventional method in early detection of tumor. The cellular quantitative parameters, such as SPF, PI and 5c Ex, are able to guide clinical work, specially in diagnosing of head and neck tumor, estimating biological character of malignant tumor and evaluating the area of excision.

Cell Nucleus↗

Early diagnosis of mesothelioma in serous effusions using AgNOR analysis.

OBJECTIVE: To compare the results of conventional cytology, DNA image cytometry, immunocytochemistry and argyrophilic nucleolar organizer region (AgNOR) analysis for the diagnosis of malignant cells in serous effusions. STUDY DESIGN: One hundred twenty effusions, 40 with carcinoses, 40 with malignant mesotheliomas and 40 without tumor cells on follow-up were studied by conventional cytology and three adjunctive methods. RESULTS: Unequivocal tumor cells were detected in 92.5% of effusions due to carcinoses and in 45% due to mesotheliomas. Applying immunocytochemistry with BerEP-4 positivity and DNA image cytometry with aneuploidy as a marker revealed 100% of carcinoses and 71.7% of mesotheliomas. Applying the experimentally found thresholds of 2.5 AgNORs as "satellites" and 4.5 AgNORs as "satellites and clusters" together as mean values per nucleus resulted in a 95% correct rate of mesothelioma and 100% rate of carcinoma cell identification without false positive diagnoses. CONCLUSION: AgNOR analysis may be a useful adjunct to other methods in the routine diagnosis of malignant serous effusions. It seems to be the most sensitive method in early cytologic diagnosis of mesotheliomas in effusions. Seventy-three percent of malignant mesotheliomas were diagnosed cytologically at first on effusions. Forty-seven percent of patients with malignant mesotheliomas were identified at the early tumor stage T1 N0 M0.

Adolescent↗

Image cytometric bcl-2:bax and bcl-2:bcl-x ratios in invasive breast carcinoma: correlation with prognosis.

BACKGROUND: The bcl-2 family of proteins are important regulators of apoptosis. Some of the members, such as bcl-2 and bcl-x(L), inhibit cell death, whereas others, such as bax and bcl-x(S), promote cell death. We evaluated the ratios of bcl-2:bax and bcl-2:bcl-x expression by image cytometry in invasive breast carcinoma to determine prognostic significance. DESIGN: Five-micron sections of formalin-fixed, paraffin-embedded tissue from 88 invasive breast carcinomas were immunostained using steam antigen retrieval, an avidin biotin-complex technique with automated stainer and primary antibodies against bcl-2 (1/160; Dako, Carpenteria, CA), bax (1/1,500; PharMingen, San Diego, CA), and bcl-x (1/1,500; PharMingen). Positive controls were tonsil (bcl-2) and normal breast (bax and bcl-x) tissue samples. Immunostain was measured in 15 high power fields as percentage positive area (PPA) in nuclei and cytoplasm using the CAS 200 image analyzer (Becton Dickinson, San Jose, CA). RESULTS: Median follow-up was 105 months (range 11-130). Significantly improved disease-free survival was found in patients with a bcl-2:bcl-x ratio > or = 1 by univariate and multivariate analyses. The bcl-2:bax ratio was not predictive of overall or disease-free survival. A significant difference in overall and disease-free survival was found between carcinomas with positive and negative bcl-2 expression by univariate analysis; by multivariate analysis, bcl-2 expression was an independent prognostic factor for disease-free survival. The 5-year survival rates were 77% and 50% in patients with bcl-2-positive and bcl-2-negative carcinomas, respectively. CONCLUSION: A bcl-2:bcl-x ratio > or = 1, assessed by image cytometry, is significantly associated with improved disease-free survival in patients with invasive breast carcinoma. Significantly increased overall and disease-free survival is associated with positive bcl-2 expression.

Adult↗

Oncogenic human papillomaviruses and ploidy in cervical lesions.

AIM: To compare ploidy measurements obtained on tissue sections of selected low and high grade squamous intraepithelial lesions containing oncogenic HPV (types 16, 18 or 33) detected by in situ hybridisation (ISH) or PCR. METHODS: DNA ploidy was assessed by image cytometry after Feulgen staining of contiguous serial sections of eight lesions exhibiting atypical squamous cells or squamous atypia and 53 low and 63 high grade squamous intraepithelial lesions in which HPV had been detected by ISH or PCR. RESULTS: Aneuploidy was strongly associated with the presence of oncogenic HPV, being detected in 50% of lesions with squamous atypia and 75.5% of the low and 95.2% of the high grade squamous intraepithelial lesions. The multiploid profile was highly associated with high grade lesions and with the pattern of HPV DNA integration. CONCLUSIONS: The presence of aneuploidy is strongly suggestive of the presence of oncogenic HPV types. Combining the detection of HPV by ISH and PCR with DNA image cytometry may provide the pathologist and the physician with important prognostic information about low grade lesions, especially when these lesions have a multiploid DNA profile and contain oncogenic HPV.

Adult↗

[DNA cytometric studies of Barrett mucosa].

BACKGROUND: Barrett's esophagus predisposes to cancer development. The diagnosis of dysplastic precursor lesions is impaired by problems of subjectivity. In the present study, DNA ploidy results of patients with Barrett's esophagus were related to the grade of dysplasia and to the progress of the disease. PATIENTS AND METHODS: In 61 patients with Barrett's esophagus, DNA ploidy was determined by image cytometry on imprints. All biopsies (n = 145) were histologically examined for dysplasia distinguishing between low- and high-grade dysplasia, and dysplasia-negative. RESULTS: A significant correlation (p < 0.001) between DNA ploidy and the grade of dysplasia was found, with 81% euploid DNA results in biopsies negative for dysplasia and 86% abnormal DNA patterns in biopsies with high-grade dysplasia. The subgroup of low-grade dysplasia showed a heterogeneous DNA ploidy. It is important to note that nearly 20% of biopsies classified dysplasia-negative contained abnormal single cells or abnormal stemlines regarding DNA content. In several cases, suspect DNA results caused a modified reclassification of dysplasia, and there were cases in which patients negative for dysplasia but positive for abnormal DNA ploidy developed dysplasia later on. CONCLUSION: In patients with Barrett's esophagus, DNA ploidy by image cytometry is a suitable additive method for histological evaluation of dysplasia. It seems to be helpful in identifying patients at risk, perhaps before the development of clear dysplasia.

Aneuploidy↗

Effects of the histone deacetylase inhibitor trichostatin A on nuclear texture and c-jun gene expression in drug-sensitive and drug-resistant human H69 lung carcinoma cells.

BACKGROUND: Texture analysis of chromatin patterns by image cytometry can be used in the development and refinement of diagnosis and prognosis of cancers and in the follow-up of therapies. However, little is known about the biological mechanisms underlying these patterns. Epigenetic mechanisms as histone posttranslational modifications and particularly histone acetylation could play a major role in the determination of these chromatin patterns and then influence nuclear texture measurements. METHODS: This study examined the consequences of treatment by the histone deacetylase inhibitor trichostatin A (TSA) on the nuclear texture in human cell lines sensitive and resistant to chemotherapy. Small cell lung carcinoma H69 cells and their variant H69-VP, which is resistant to etoposide, were incubated with 100 ng/ml of TSA for 0 to 24 h. Nuclear texture was evaluated by image cytometry and compared with the histone H4 acetylation level measured by western blotting and expression of c-jun gene evaluated by reverse transcription and real-time polymerase chain reaction. RESULTS: TSA treatment induced an increase in histone H4 acetylation level in both cell lines. However, at the level of chromatin texture, sensitive H69 cells displayed a progressive chromatin decondensation up to 24 h, whereas resistant H69-VP showed rapid (8 h) but transient changes. Similarly, expression of c-jun increased regularly in TSA-treated H69 cells. In H69-VP cells, an increase was also observed up to 12 h followed by a decrease after 24 h of treatment. CONCLUSIONS: Analysis of nuclear texture appeared to be a sensitive technique to detect chromatin pattern alterations induced by the histone deacetylase inhibitor TSA in the H69 cell line and enabled the observation of chromatin pattern discrepancies between chemotherapeutic drug-sensitive and drug-resistant cells during this treatment. When c-jun gene expression was analyzed as gene sensitive to epigenetic control, these textural differences seemed to be correlated to gene expression.

Carcinoma, Small Cell↗

Near tetraploid prostate carcinoma. Methodologic and prognostic aspects.

BACKGROUND: The clinical value of DNA ploidy analysis in prostate carcinoma has been an issue for investigation for more than 2 decades. In general, diploid or pseudodiploid tumors are associated with a favorable prognosis and aneuploid tumors with an unfavorable prognosis, irrespective of type of treatment. Tumors with DNA values in the tetraploid region (around 4c) present a diagnostic problem. Such DNA distributions may clearly represent aneuploid tumors with an unfavorable prognosis. However, a 4c distribution may conversely represent a tetraploid tumor (possibly a polyploid variant of the diploid tumor) with a favorable prognosis. Previous data from our laboratory indicate the existence of such a tetraploid subgroup. The goal of the current study was to investigate the diagnostic problem of 4c tumors in greater detail. METHODS: Ploidy classification of cytologic smears by image cytometry was performed in a retrospective study of 334 patients with hormonally treated prostate carcinoma. Follow-up time was 30 years or until death. RESULTS: Three ploidy types were defined: near-diploid (D type), near-tetraploid (T type), and highly aneuploid (A type). Tumors with a modal value within the tetraploid region were found in 27% (92 cases) of the total material. Of these, 9% were defined as T type and 18% as A type. Overall, 37% of the tumors were classified as D type, 9% as T type, and 54% as A type. Of the A type tumors, one-third had modal DNA values in the tetraploid (4c) region. Multivariate analysis showed a statistically significant difference between A type tumors and D and T type, but not between D type and T type. Both D and T type tumors progressed slowly and killed the patients 5 to 30 years after diagnosis, whereas A type tumors progressed rapidly and killed the patients within 6 years of diagnosis. CONCLUSIONS: By image cytometry, prostate carcinoma can be divided into three ploidy types: D, T, and A type. Biologically, however, the tumors fall into only two groups: low grade malignant, pseudodiploid tumors of D or T type, and high grade malignant, highly aneuploid tumors of A type.

Carcinoma↗

Large-scale genomic instability predicts long-term outcome for women with invasive stage I ovarian cancer.

BACKGROUND: The objective was to evaluate the value of DNA ploidy using high-resolution image cytometry in predicting long-term survival of patients with early ovarian cancer. PATIENTS AND METHODS: A retrospective analysis of 284 cases with FIGO stage I ovarian carcinoma treated during the period 1982-1989 was performed. Clinical follow-up information was available for all patients. RESULTS: Patients with diploid and tetraploid tumors had a 10-year relapse-free survival of 95% and 89%, respectively, compared with 70% and 29% for polyploid and aneuploid tumors, respectively. DNA ploidy analysis was the strongest predictor of survival in multivariate analysis (diploid/tetraploid versus polyploid/aneuploid; relative hazard 9.0) followed by histological grade, including clear cell tumors in the group of poorly differentiated tumors (grade 1-2 versus grade 3 or clear cell; relative hazard 2.7), and FIGO stage (Ib/Ic versus Ia; relative hazard 2.0). In a stratified Kaplan-Meier analysis, patients with grade 1-2, diploid or tetraploid tumors had a 10-year relapse-free survival of 95%, forming a low-risk group. Patients with grade 3 or clear cell, diploid or tetraploid tumors had 10-year relapse-free survival of 86%, forming an intermediate-risk group, while all patients with aneuploid/polyploid tumors formed a high-risk group, with 10-year relapse-free survival of 34%. CONCLUSIONS: This study points to the importance of including DNA ploidy analysis by image cytometry when selecting patients with early ovarian cancer for adjuvant treatment after surgery.

Adult↗

[DNA synthesis in the testes of infertile men with varicocele--image cytometeric analysis of proliferating cell nuclear antigen].

BACKGROUND: The analysis of cell kinetics of germ cells have been implicated as important clue to investigate the etiology of infertile patients. We examined the germ cell kinetics of varicocele testis using proliferating cell nuclear antigen (PCNA) immunostaing by image cytometry (CAS 2000), which is very accurate and useful method to evaluate the staining intensity. PATIENTS AND METHODS: Testicular biopsy specimen obtained from 28 cases of male infertility with left varicocele and 5 normal controls were quantitatively examined concerning PCNA expression. Positive rate of PCNA (PR) was expressed as the percent of nuclear area positively stained with PCNA monoclonal antibody compared with total nuclear area analyzed in more than 20 seminiferous tubules. A hematoxylin and eosin-stained sections, which were fixed in Bouin's solution were used to examine Johnsen's mean score by photomicroscopy. RESULTS: PR of right and left testes of varicocele patients were 11.3 +/- 4.8% and 11.3 +/- 4.1%, respectively. PR of normal testes was 21.7 +/- 3.1%. PR of each side were reduced similarly and the difference of PR between right and left testes of varicocele patients were not significant. Bilateral PR was 11.4 +/- 4.1%. Bilateral PR was significant lower than that of normal testes (p < 0.001). Concerning varicocele grade, sperm concentration of seminogram, Johnsen's mean score and the improvement of seminogram, no significant correlations between PR and those were observed. CONCLUSION: PR evaluated by image cytometry is very useful biological parameter to assess the germ cell kinetics. In varicocele patients, not only left side but also right side of testes were deteriorated in the cell kinetics and the decrease in DNA synthesis is one of the causes that have deleterious effects on spermatogenesis.

Adult↗

Diagnostic value of DNA analysis in effusions by flow cytometry and image analysis. A prospective study on 102 patients as compared with cytologic examination.

One hundred twenty-six effusion samples from 102 patients were examined by cytology and flow cytometry (FCM). Overall, there was an 84% correlation between cytologic and FCM results. Of the 36 malignant cases determined by cytologic examination, FCM revealed an aneuploid peak in 20 (56%). Image analysis (IA) performed on the malignant cytologic cases with a diploid flow pattern detected two additional aneuploid peaks. In addition, FCM indicated three aneuploid cases in which cytologic characteristics were initially interpreted as benign (false negative). Aneuploidy was therefore detected in 64% of the malignant effusion specimens by FCM and IA. Twenty-three of the total of 24 aneuploid cases detected by FCM were associated with malignancy (predictive value = 96%). The one nonmalignant case was that of hemorrhagic pancreatitis with infected pseudocyst. FCM is an excellent tool when moderate to large numbers of tumor cells are present, whereas use of IA is advantageous for specimens containing smaller numbers of malignant cells because these can be directly analyzed. When an aneuploid peak is present, a diagnosis of malignancy must be suspected, and, if the initial cytologic screen is negative, a critical review of the cytology slides is justified. In those cases with an equivocal atypical cytology report and an abnormal cytometric histogram, additional investigation is warranted. In some malignancies the tumor cells will be diploid (in this study 36%) and neither FCM nor IA will add to tumor detection, leaving cytologic examination as the definitive technique.

Adult↗