Image analysis combined with quantitative cytochemistry

Ploem, J. S.; Driel-Kulker, A. M. J. van; L, Goyarts-Veldstra; Ploem-Zaaijer, J. J.; Verwoerd, N. P.; Zwan, Matthew van der

doi:10.1007/bf00482990

Cited by 27 publications

(9 citation statements)

References 9 publications

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“…For automated detection and quantification of rareevent cells, two main approaches can be distinguished: flow cytometry (Herzenberg et al 1979;Lewis et al 1996) and image cytometry (Ploem et al 1986;Mesker et al 1994;Tanke et al 1996). Both methods rely heavily on the use of highly specific markers for cell recognition.…”

Section: The Concept Of Rare-event Detectionmentioning

confidence: 99%

Strategies for Rare-Event Detection: An Approach for Automated Fetal Cell Detection in Maternal Blood

Oosterwijk¹,

Knepflé²,

Mesker³

et al. 1998

The American Journal of Human Genetics

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This article explores the feasibility of the use of automated microscopy and image analysis to detect the presence of rare fetal nucleated red blood cells (NRBCs) circulating in maternal blood. The rationales for enrichment and for automated image analysis for "rare-event" detection are reviewed. We also describe the application of automated image analysis to 42 maternal blood samples, using a protocol consisting of one-step enrichment followed by immunocytochemical staining for fetal hemoglobin (HbF) and FISH for X- and Y-chromosomal sequences. Automated image analysis consisted of multimode microscopy and subsequent visual evaluation of image memories containing the selected objects. The FISH results were compared with the results of conventional karyotyping of the chorionic villi. By use of manual screening, 43% of the slides were found to be positive (>=1 NRBC), with a mean number of 11 NRBCs (range 1-40). By automated microscopy, 52% were positive, with on average 17 NRBCs (range 1-111). There was a good correlation between both manual and automated screening, but the NRBC yield from automated image analysis was found to be superior to that from manual screening (P=.0443), particularly when the NRBC count was >15. Seven (64%) of 11 XY fetuses were correctly diagnosed by FISH analysis of automatically detected cells, and all discrepancies were restricted to the lower cell-count range. We believe that automated microscopy and image analysis reduce the screening workload, are more sensitive than manual evaluation, and can be used to detect rare HbF-containing NRBCs in maternal blood.

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Section: The Concept Of Rare-event Detectionmentioning

confidence: 99%

Strategies for Rare-Event Detection: An Approach for Automated Fetal Cell Detection in Maternal Blood

Oosterwijk¹,

Knepflé²,

Mesker³

et al. 1998

The American Journal of Human Genetics

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“…However, they offer the advantage that an image of each detected event can be stored in memory for visual evaluation afterwards. When necessary, detected cells can also be relocated using the microscope, because the X,Y coordinates of each detected object are known (15,16). This advantage has made image cytometry very well suited to detect rare events in frequencies below 1 in lo5, a detection level that is generally not achievable by flow cytometry (5,6).…”

mentioning

confidence: 99%

Detection of immunocytochemically stained rare events using image analysis

et al. 1994

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The detection of rare-event cells circulating in peripheral blood using automated image analysis was evaluated using a model system consisting of cells from a breast cancer cell line (SKBR3) seeded in a mononuclear cell suspension. Slides of cells with optimal morphology were prepared according to an optimized preparation procedure based on centrifugal cytology in combination with formalin fixation.SKBR3 cells were immunocytochemically stained for cytokeratin using the cam 5.2 monoclonal antibody and labelled with alkaline phosphatase using CAS-red as substrate. Because, for optimal segmentation of cell images, plain differences in absorption wavelength are required, the red immunostaining was combined with a green nuclear counterstaining based on ethyl green. Slides were automatically screened for cytokeratin-positive SKBR3 cells resulting in a lowest detectable frequency of one positive cell per 1.87 x lo6 negative cells. A comparison between manual screening and automated screening for cytokeratin-positive cells showed a high level of correlation (0.9998).For the definition of the total number of objects per slide, two counting procedures were evaluated. Results were close to the visual score with a coefficient of variation of 0.47% for the counting procedure used in this study.It is concluded that optimization of preparation and staining procedures for the detection of rare-event cells using automated image analysis results in optimal image contrast and, consequently, in an increase in sensitivity for detecting rare events. 0 1994 Wiley-Liss, Inc.

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“…Staining techniques have also been evaluated and standardized for the requirements of automation (Ploem et al, 1986;Wittekind, 1985).…”

Section: Stainingmentioning

confidence: 99%

“…The influence of routine staining techniques on the efficiency of segmentation and feature extraction was an early consideration in work aimed at automated cytological analysis (Holmquist et al, 1976;Wittekind et al, 1983). Staining techniques have also been evaluated and standardized for the requirements of automation (Ploem et al, 1986;Wittekind, 1985).…”

Section: Stainingmentioning

confidence: 99%

The microcomputer and image analysis in diagnostic pathology

Jarvis

1992

Microscopy Res & Technique

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This paper presents a snapshot view of the influence and direction of microcomputer technology for image analysis techniques in diagnostic pathology. Microcomputers have had considerable impact in bringing image analysis to wider application. Semi-automated tracing techniques are a simple means of providing objective data and assist in a wide range of diagnostic problems. From the common theme of reducing subjectivity in diagnostic assessment, an extensive body of research has accrued. Some studies have addressed the need for quality control for reliable, routine application. Video digitizer cards bring digital image analysis within the reach of laboratory budgets, providing powerful tools for investigation of a wide range of cellular and tissue features. The use of staining procedures compatible with quantitative evaluation has become equally important. As well as assisting scene segmentation, cytochemical and immunochemical staining techniques relate the data to biological processes. With the present state of the art, practical use of microcomputer based image analysis is impaired by limitations of information extraction and specimen throughput. Recent advances in colour video imaging provide an extra dimension in the analysis of multi-spectral stains. Improvements will also be felt with predictable increase in speed of microprocessors, and with single chip devices which deliver video rate processing. If the full potential of this hardware is realized, high-speed, routine analysis becomes feasible. In addition, a microcomputer imaging system can play host to companion functions, such as image archiving and transmission. With this outlook, the use of microcomputers for image analysis in diagnostic pathology is certain to increase.(ABSTRACT TRUNCATED AT 250 WORDS)

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Image analysis combined with quantitative cytochemistry

Cited by 27 publications

References 9 publications

Strategies for Rare-Event Detection: An Approach for Automated Fetal Cell Detection in Maternal Blood

Strategies for Rare-Event Detection: An Approach for Automated Fetal Cell Detection in Maternal Blood

Detection of immunocytochemically stained rare events using image analysis

The microcomputer and image analysis in diagnostic pathology

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