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At least 19 recordsLinked to original sources

PubMed1985

A charge coupled device-based image cytophotometry system for quantitative histochemistry and cytochemistry.

A rapid, semiautomated cytophotometry system for quantitative histochemistry and cytochemistry was constructed. The system consists of a Fairchild charge coupled device (CCD) image camera, a Zeiss Universal microscope, a Datacube analog to digital converter, and a digital Equipment Corporation LSI 11/23 computer operating under RT-11. Computer programs were written in FORTRAN and the MACRO assembly language for the acquisition of data from the CCD device. These data were then used for image segmentation, image display, and calculation of total optical density, perimeter, cell area, and several shape features. The reproducibility of measurement made with the CCD-based cytophotometry system was tested by repeated measurements. The coefficient of variation was estimated to be 1.7% for total optical density and 0.9% for cell area. The CCD-based cytophotometry system was further evaluated by comparing results with measurements made on the same cells with a scanning stage cytophotometer using the HIDACSYS computer programs. Correlation coefficients of 0.96 for total optical density and 0.91 for cell area were obtained between the two systems. We conclude that the high-speed, dimensional stability, small size, and linearity of the CCD-based cytophotometry system will make it useful for quantitative histochemistry and cytochemistry.

Animals↗

PubMed1988

DNA cytophotometry in malignant thyroid tumors--use of different evaluation schemes for prognostic statements.

Quantitative evaluation of nuclear DNA has been found to provide information on diagnosis and prognosis in a number of malignant tumours and borderline lesions. Using 53 carcinomas of the thyroid with varying differentiation we examined the prognostic information obtained by DNA cytophotometry with respect to clinical outcome, applying three different evaluation schemes. DNA cytophotometry allowed the discrimination of euploid carcinomas with good prognosis from aneuploid tumours with bad prognosis and a generally shortened life span. However, the encapsulated variants of follicular and papillary thyroid carcinomas, with their generally excellent prognosis, exhibited DNA histograms similar to those of their widely invasive counterparts. Thus, the favourable prognosis of these minimal invasive subtypes may primarily be related to the tumours' encapsulation and cannot be ascribed to a particular DNA content. We conclude that DNA cytophotometry can provide additional prognostic information for the individual patient suffering from thyroid carcinoma and may probably lead to an individualization of the therapeutic strategies.

Adenocarcinoma↗

PubMed1988

Retrospective DNa analysis of T3/T4 breast carcinoma using cytophotometry and flow cytometry. A comparative study with prognostic evaluation.

Tumor cells from 72 patients with advanced breast carcinoma (T3/T4) were analyzed for their DNA content by cytophotometry and flow cytometry (FCM). Both methods were able to subdivide the tumors into groups with different prognoses. Patients with a normal/near-normal DNa content in the tumor had a better prognosis than did those with aneuploid tumors. FCM measurements of DNA content gave a better discrimination for both survival (P = .019) and disease-free survival (P = .059) than did cytophotometry (P = .105 and P = .067, respectively). These results indicate that FCM analysis of paraffin-embedded material is a more accurate method than is cytophotometry for characterizing the DNA content of breast carcinoma cells. The method could be useful in both retrospective studies and in daily diagnostic work.

Adult↗

PubMed1978

Comparison of red and white muscles by cytophotometry of their muscle fibre populations.

Samples from two red muscles (vastus intermedius and vastus medialis) and two white muscles (biceps femoris and gluteus medius) were taken from four pigs. Serial transverse sections were reacted for ATPase and NADH oxidative activity. Sections were mapped with a projection microscope so that the staining intensity of individual fibres for the two reactions could be measured with a simple microscope photometer. Transmission values at 600 nm were converted to units of 0-10 for the range from darkest to lightest staining fibres on each section to cancel variation in staining intensity between sections. The aim of the study was to use simple cytophotometry instead of subjective judgement in the categorization of different histochemical types of muscle fibres. Cytophotometry enabled clear resolution of the major fibre types (types I and II using the ATPase reaction), partial resolution of more variable characteristics (NADH oxidative activity in type I and II fibres) and no resolution of subtle subtypes (IA and IB with the NADH oxidative reaction). However, between the major fibre types, cytophotometry revealed variable numbers of fibres with transitional characteristics. There were more of these fibres in red muscles. With sections reacted for ATPase, transmission values for low magnification fields containing 100 to 200 fibres were correlated (r = -0.91) with the ratio of type I:II fibres.

Adenosine Triphosphatases↗

PubMed1987

Flow cytometry and Feulgen cytophotometry in evaluation of effusions.

Fifty-eight effusions (42 pleural and 16 ascitic fluids) from patients with and without cancer were analyzed by conventional cytology and the results compared with DNA patterns generated by flow cytometry of 10(4) nuclei and several modes of Feulgen cytophotometry. In 31 patients (24 without evidence of cancer and seven with history of cancer and cytologically negative fluids), the fluids were diploid by flow cytometry. One fluid with atypical cells from a lymphoma suspect was also diploid. Flow cytometry of 26 cytologically cancerous fluids disclosed aneuploid DNA patterns in 16 and diploid patterns in ten. Feulgen cytophotometry of 11 of these fluids (three aneuploid, eight diploid) was performed on nuclear preparations identical to those used in flow cytometry and on restrained smears used for visual analysis. The analysis was performed in two modes: as a study of 500 sequential nuclei in an automated system, mimicking flow cytometry, and visually selected large, presumably malignant nuclei. In nine of the 11 cases, the DNA content of visually selected cancer cells was aneuploid, even though this DNA pattern was not evident in the analysis of 500 sequential cells. In two cases, both diploid by flow cytometry, the Feulgen analysis confirmed the presence of cancer cells in the diploid range. In samples of 10(4) nuclei representing a mixed population of cells occurring in effusions, the presence of aneuploid cancer cells may not be disclosed by conventional flow cytometry. A larger sample of cells, a detailed analysis of DNA histograms, and perhaps sorting of select cells in the hypertetraploid range, may prove essential before flow cytometry can be accepted as a diagnostic tool in the laboratory in the assessment of effusions.

Aneuploidy↗

PubMed1992

DNA cytophotometry and prognosis in ovarian tumors of borderline malignancy. A clinicomorphologic study of 80 cases.

Surgical specimens of 80 ovarian tumors of borderline malignancy (OTBM) were investigated by scanning DNA cytophotometry. Diploid or euploid DNA histograms were found for 21 tumors, whereas 59 OTBM showed noneuploid or aneuploid DNA patterns. All patients were followed-up after surgery for at least 3 years (mean observation period, 6.7 years). Follow-up showed 11 cases of recurrent disease and 6 deaths. DNA findings and several other morphologic and clinical details (including patient age, histologic type and stage of disease, and extent of therapy) were correlated to the postoperative course. Statistical analyses disclosed that, of these parameters, only DNA content significantly affected prognosis. Recurrences and deaths resulting from tumor exclusively were observed among patients with noneuploid or aneuploid OTBM, whereas none of the diploid or euploid tumors recurred (P less than 0.05). DNA cytophotometry thus might be regarded as an effective complementary means to assess the prognosis of individual OTBM cases.

Adult↗

PubMed1987

Melanoma in situ (MIS) adjacent to an invasive nodular melanoma ("SSM/NM") and its metastases--DNA-cytophotometry, mitotic index, and anisokaryosis.

Malignant melanomas of the superficial spreading type usually have an intraepidermal tumour component in their periphery which frequently displays the morphological features of a melanoma in situ (adjacent MIS). It is thus comparable to exclusively epidermal melanomas; melanoma in situ (MIS). Taking 10 superficial melanomas with a nodular component ("SSM/NM") 31 adjacent MIS regions and 36 nodular melanoma components were analysed in serial tissue slides. Planimetric estimation of the nuclear areas was employed as a measure of anisokaryosis. DNA-Feulgen-cytophotometry was applied to obtain an objective variable in judging malignancy in the DNA-histographs (paraffin material). Furthermore we investigated 8 metastases of one of the malignant melanomas applying the methods described. A comparison of the epidermal with the invasive tumour components revealed an increase in the nuclear area which, however, decrease from the superior to the inferior nodular regions and which are further reduced in melanoma metastases. Anisokaryosis is evidently less in metastases compared with all primary melanomas. The nuclear DNA-content increases from the epidermal to the invasive tumour compartments and is lower in the inferior nodular regions when compared with the superior ones. No further significant differences are, however, established in the metastases. The coefficients of variability of the DNA-contents, being a potential indicator of DNA-heterogeneity reflect higher values in the epidermal tumour components compared with the nodular regions, decreasing from the superior to the inferior nodular parts of the tumour. All metastases have smaller values than the respective primary melanoma. In the DNA-histographs 75% of the intraepidermal tumour components have obvious signs of malignancy including tumour cell stem lines in 19% of the cases. 85% of the nodular regions investigated have clear signs of malignancy, 33% of which also have aneuploid stem lines. All metastases have obvious signs of malignancy and tumour cell stem lines in 50% of the cases observed. The following conclusions can be drawn from our findings: DNA-Feulgen-cytophotometry and nuclear planimetry are additional feasible methods for judging the epidermal component of a melanocytic lesion as malignant (adjacent melanoma in situ) on paraffin material. Furthermore these methods give different results in invasive nodular versus epidermal (in situ) melanoma components. Both the DNA-histographs and our immunohistochemical investigations (monoclonal antibody P 3.58) indicate the malignant potential of adjacent MIS.(ABSTRACT TRUNCATED AT 400 WORDS)

Cell Nucleus↗

PubMed1990

Cytophotometry in tumor pathology. A critical review of methods and applications, and some results of DNA analysis.

In tumor pathology the quantitation of cellular substances can be of diagnostic value. Microscope cytophotometry and digital image analysis and, on the other hand, flow cytometry are supplementary methods for measuring, each with a typical spectrum of application. The methods are predominantly used for DNA analysis: Static and image cytophotometry are applicable to cytologic and histologic slides preferably for identifying stem lines in tumors of heterogenous morphology and in merely circumscribed lesions (e.g., precancerous lesions). On the other hand, sampling errors due to preselection, and the often low number of cells actually measured, may preclude the possibility of exact cell cycle analysis. This is, in fact, an important additional option of flow cytometry resulting from the high resolution of DNA histograms, which is explained by the large number of cells that can be measured in a short period. Sampling errors in flow cytometry may result from the preparation of single cell suspensions which in certain tumor entities may suppress a varying amount of particularly fragile cells or nuclei. The prognostic significance of DNA ploidy, stem line heterogeneity and S-phase fraction is clearly described in quite a number of tumor entities. Independent of its prognostic value, the cytometric identification of stem lines might be particularly useful in the follow-up of tumor patients, where it may indicate the effectivity of systemic therapy. The development of therapeutic concepts is aptly supported by flow cytometric cell cycle analysis which helps to assess the in vitro effect of combined cytostatics on the proliferative process. Moreover, multiparameter analysis of biopsy samples may provide greater accuracy in characterising individual tumor stem lines and may furthermore help to develop improved protocols for the therapy of solid tumors.

Cytophotometry↗

PubMed1988

DNA-cytophotometry of lymph node touch imprints in cutaneous T-cell lymphoma.

Scanning DNA-cytophotometry was performed on touch imprints of 26 lymph nodes (LN) obtained from 25 patients with cutaneous T-cell lymphoma (CTCL), stained by the Feulgen technique, and interpreted without knowledge of histopathologic diagnosis. Four patterns of DNA distribution were identified, but only histograms that demonstrated cells containing nuclei with more than 4C DNA content (hypertetraploidy) reliably distinguished LN involved with CTCL from LN with reactive changes; for example, dermatopathic lymphadenitis. An abnormal DNA histogram with evidence of hypertetraploidy was demonstrated in 9 of 12 LN showing histopathologic evidence of involvement compared with no abnormal histograms in 14 LN without histopathologic involvement. One LN that was diffusely involved with CTCL had a DNA distribution characteristic of a relatively high level of cell proliferation, but without definite hypertetraploidy. Cytogenetic studies on the blood of this patient, who had Sézary syndrome, demonstrated a clone of lymphocytes with a pseudodiploid karyotype without a related polyploid subline. The remaining two histopathologically involved LN had normal DNA histograms; these LN were only focally involved with CTCL. These observations indicate that DNA-cytophotometry correlates well with the histopathologic findings in LN diffusely involved with CTCL, but may be normal in LN with focal involvement or in those that contain cytogenetically abnormal cells with a near-diploid DNA content.

Adult↗

PubMed1985

[Morphology and growth behavior of ovarian tumors. Studies using DNA cytophotometry, immunohistology and tissue culture].

Antigens A, B, Lea, Leb, CA 12-5, CA 19-9 and CEA were prepared immunohistologically, a DNA-cytophotometry was carried out and the value of a new in vitro proliferation assay to test cytostatic drugs investigated. Material was as follows: 20 normal ovaries and 105 ovarian tumors. Blood group antigens were expressed at a high percentage from well-differentiated epithelial cells, CEA in 50% and CA 12-5 in 70% of the ovarian carcinoma. Predictions about the dignity of borderline tumors can be made with DNA-cytophotometry. The efficacy of 6 different cytostatic agents was investigated in a new in vitro proliferation assay.

Antigens, Neoplasm↗

PubMed1988

In situ detection of human immunodeficiency virus (HIV) nucleic acid in H9 cells using nonradioactive DNA probes and an image cytophotometry system.

Rapid and sensitive nonradioactive methods to detect human immunodeficiency virus (HIV)-infected cells are needed in clinical medicine. We developed an in situ hybridization test using 2-acetylaminofluorene (AAF)-labeled HIV DNA as a hybridization probe. Hybridized probe was detected using rabbit anti-AAF antibody, followed by alkaline phosphatase-conjugated goat anti-rabbit, and the bromochloroindolyl phosphate-nitroblue tetrazolium reaction. An image cytophotometry system was used to quantitate the percentage of HIV-infected cells. These methods were used to determine the percentage of H9 cells infected with HIV. HIV was detected in 0% of cells on day 1 post infection, 7% on day 4, 41% on day 8, and 5% on day 15. These results paralleled those of the reverse transcriptase assay and an antigen capture ELISA assay for HIV antigen. Thus the AAF modified HIV DNA probe detected HIV nucleic acid in infected H9 cells and the image cytophotometry system improved the sensitivity and objectivity of detection.

2-Acetylaminofluorene↗

PubMed1990

Single cell DNA cytophotometry in clinical stage I malignant melanoma. Relationship to prognosis.

Histological sections of 50 clinical stage I cutaneous melanomas were analyzed by single cell DNA cytophotometry. Forty-two percent of the melanomas had diploid modal values. Ploidy was not significantly related to the level of invasion, melanoma thickness or prognosis. These results are contradictory to published data from flow cytophotometry which, however, differ inter alia concerning patient materials and follow-up times. Mean nuclear area was in our study significantly correlated to the prognosis.

Adult↗

PubMedDate not supplied

Absorption cytophotometry: evaluation of three methods for the determination of DNA in Feulgen stained nuclei.

Three methods for the evaluation of the relative amount of DNA per nucleus by absorption cytophotometry are compared. A combination of the two-wavelength method (Patau, 1952; Ornstein, 1952), the one-wavelength two-area method (Garcia, 1965) and the determination of transmission through nuclear plugs is proposed in order to estimate systematic errors made by absorption cytophotometry.

DNA↗

PubMed1986

The application of flow cytophotometry in measurements of cell adhesion.

A common approach to the study of cell substrate interactions is the measurement of the attachment of cells to different substrates or to cultured cell layers. The evaluation of attachment is made either by scintillation counting of previously labelled adhering cells, or by light microscopy using the criterion of cell shape, sometimes refined by automatic image analysis. These methods have many drawbacks. This paper suggests the use of fluorescence-activated flow cytophotometry, (FC) which yields direct counts of the non-adhering cells. These "free" cells are removed after completion of the adhesion experiment from the microtitre plate wells. An internal standard, in the form of fluorescent polystyrene beads is added, allowing evaluation of the percentage of cells adhering to the well walls. Flow cytophotometry then produces data based on the analysis of large populations of cells. Unequivocal discrimination is obtained between the counted cells and counted fluorescent beads eliminating counting errors. The results can be processed on line by computer. A suspension of mouse splenocytes was used for the evaluation of the overall error of the method arising from inaccuracies in pipetting, interference of glutaraldehyde with ethidium bromide (EB) staining and instrumental error. Each adhesion experiment was terminated by staining and post-fixation and it was established that this introduces no change in cell counting, in comparison with the original unfixed cells. Prefixation, however, quenches the EB staining and would interfere with the counting procedure. The overall standard error of the technique was found to be 5%-10%.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

DNA cytophotometry in adrenocortical tumours: a clinicomorphological study of 66 cases.

Surgical specimens of 66 adrenocortical tumours were investigated by conventional microscopy and DNA cytophotometry. Histologically, 50 neoplasms were classified as adenomas and 16 as carcinomas. In only 8 of the latter cases were distant metastases and/or a lethal outcome recorded. On single cell scanning cytophotometry either non-euploid or aneuploid DNA histograms were identified in 24 of 50 adenomas (48%) and in 14 of 16 carcinomas (88%). The two carcinomas exhibiting euploid DNA distributions fell into the group of 7 malignancies which are recurrence-free so far. From these findings it is concluded that DNA measurements have no diagnostic and only limited prognostic value in neoplasms of the adrenal cortex.

Adenoma↗

PubMed1976

The significance of DNA flow-through fluorescence cytophotometry for the diagnosis of prostate carcinoma.

Flow-through fluorescence cytophotometric determination of nuclear DNA content was employed for the diagnosis of prostate carcinoma. Fine needle aspiration biopsy material from the prostate of 220 patients was used for study. A false negative rate of 11.4% and a false positive rate of 29.7% were obtained when the results of flow-through photometry were compared with those of traditional cytodiagnosis. It was found that 4.5% of the specimens were unsuitable for cytologic diagnosis and 10.9% for flow-through cytophotometry. False negative DNA histograms may be due to two factors: either the number of tumor cells is small or there are tumor cells whose nuclear DNA content does not differ from that of a normal cell population. False positive findings result from proliferating cells in inflammatory activation. Errors in preparation of the material and mechanical mistakes, such as cellular clumping and coincidences, are less likely causes. The greater percentage of specimens which were inadequate for cytophotometry was due to the large number of cells needed for a utilizable flow-through photometric histogram. The high rate of false negative and false positive results (11.4% and 29.7%, respectively) argues against using flow-through photometric nuclear DNA determination for the diagnosis of prostate carcinoma.

Biopsy, Needle↗

PubMed1979

DNA-cytophotometry of lymph node imprints from patients with mycosis fungoides.

To obtain objective criteria for early diagnosis of lymph node involvement in patients with mycosis fundoides, Feulgen DNA-cytophotometry was carried out in lymph node imprints from patients with mycosis fungoides. The lymph nodes of 3 patients with lymph nodes, showing partial or total replacement by atypical lymphoreticular tissue histologically, showed a polyploid and aneuploid DNA distribution. Eleven out of 22 patients with dermatopathic lymphadenopathy both with or without early involvement histologically had an abnormal DNA histogram with hypertetraploid DNA values. Four of these 11 died, 5 had a partial remission in response to therapy and 2 had sustained remission during the follow-up period of 5 yr. The other 11 patients had a normal DNA distribution. Of these 11, one died and 10 achieved sustained remission after therapy. There is a good correlation between the DNA-cytophotometric results and histology of the lymph nodes. On the basis of these results DNA-cytophotometry may be considered an additional and objective aid in the diagnosis of lymph node involvement in mycosis fungoides.

Adult↗

PubMed1977

Studies on the population kinetics of the Walker carcinoma by autoradiography and pulse cytophotometry.

The proliferation parameters of the Walker carcinoma were estimated from both in vivo and in vitro measurements. tthe transplantable Walker carcinoma 256 was grown in male inbred BD1 rats. During exponential growth, 5--6 days after transplantation, a PLM curve was performed, yielding estimates of TC approximately equal to 18-0 hr, TS approximately equal to 6-4 hr, TG2+M approximately equal to 4-1 hr. With the double labelling technique in vitro under 2-2 atm oxygen we obtained: TC approximately equal to 18-2 hr, TS approximately equal to 8-2 hr, TG2+M approximately equal to 2-0 hr. From pulse cytophotometry DNA content histograms the fractions of cells in the cell cycle phases were calculated using a computer program: fG1 approximately equal to (47-6 +/- 1-1)%, fS approximately equal to (34-1 +/- 1-0)%, fG2+M approximately equal to (18-3 +/- 1-5)%. These fractions remained constant between the fifth and the twelfth day after transplantation. At that time the tumour growth had already slowed down appreciably. The growth fraction determined by repetitive labelling was 0.96 on the fifth and 0-93 on the seventh and eleventh day. The cell loss factor was phi approximately equal to 17% during exponential tumor growth and increased to about 100% between the tenth and twelfth day. The agreement of the cell kinetic data determined by autoradiography from solid tumours in vivo (PLM, continuous labelling) and autoradiography as well as pulse cytophotometry from in vitro experiments (excised material) was satisfactory.

Animals↗