Determination of ploidy and steroid receptor status in breast cancer by laser scanning cytometry

Bollmann, R.; Torka, Robert; Schmitz, Jörg; Bollmann, Magdolna; Méhes, Gábor

doi:10.1002/cyto.10093

Cited by 9 publications

(7 citation statements)

References 21 publications

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“…Due to the high number of cells and other technical reasons (e.g., the use of PI staining and the presence of microorganisms), the accuracy of the measurement was inferior compared with the results from interactive image cytometry (e.g., an increase in CV). However, we found that LSC allows a good estimation of total nuclear DNA content, as described recently 26. In the current study, LSC was used to search for rare events in cervical cytologic samples.…”

Section: Discussionsupporting

confidence: 63%

Determination of features indicating progression in atypical squamous cells with undetermined significance

Bollmann

Méhes

Torka³

et al. 2002

Cancer

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BACKGROUND.The Bethesda System of cervical cytologic findings introduced the term ASCUS (atypical squamous cells of undetermined significance) to cover the broad zone separating normal cytomorphology from definitive squamous intraepithelial lesions (SILs ). The management of patients with ASCUS is particularly problematic as approximately 10% of ASCUS patients develop SIL and 1 per 1000 develop cervical carcinoma. METHODS.Our aim was to demonstrate the combined use of polymerase chain reaction for human papillomavirus (HPV) typing and laser scanning cytometry for DNA content measurements in the subcategorization of ASCUS cases according to the risk for progression toward cancer. Liquid-based monolayer preparation (ThinPrep, Cytyc, Boston, MA) of the cytologic material was used for cytomorphologic analysis. DNA content measurements using laser scanning cytometry and direct sequencing of HPV using the consensus primers GP5ϩ/GP6ϩ and MY09/ MY11 were performed from the same material. RESULTS.Twelve of the 44 cases (27.2%) with ASCUS carried a high-risk HPV genome whereas only 3 of the 195 normal control cases (1.5%) showed positivity for a high-risk HPV genome. Six of 12 (50%) of the high-risk HPV-positive ASCUS cases presented isolated cells with a DNA content above 5c, whereas cells with a DNA content above 9c were found in 3 of 12 cases (25%) and were exclusively found in combination with high-risk HPV infection. In these three cases, the histologic follow-up resulted in cervical intraepithelial neoplasia (CIN) I (one case) and CIN III (two cases). None of the other ASCUS or normal cases displayed DNA aneuploidy above 9c. They returned to normal cytology (within normal limits/ benign cellular changes) in the follow-up. CONCLUSIONS.Human papillomavirus typing and DNA content measurements may delineate a distinct group of ASCUS. Our preliminary data suggest that ASCUS cases with high-risk HPV positivity and with rare cells with abnormally high DNA content represent similar biologic features as high-grade SIL and are at elevated

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Section: Discussionsupporting

confidence: 63%

Determination of features indicating progression in atypical squamous cells with undetermined significance

Bollmann

Méhes

Torka³

et al. 2002

Cancer

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“…The specimens can be destained and restained (68) to measure additional attributes of the same cells; the relocation feature of LSC allows one to precisely identify each cell by its location on the slide. Several publications describe the usefulness of LSC in analysis of tissue sections, FNA samples, or touch preparations (44,(73)(74)(75)(76)(77)(78)(79)(80)(81)(82)(83).…”

Section: Lsc In Clinical Pathologymentioning

confidence: 99%

Laser Scanning Cytometry

Pożarowski

Holden²,

Darżynkiewicz

2006

Cell Imaging Techniques

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The laser scanning cytometer (LSC) is the microscope-based cytofluorometer that offers a plethora of analytical capabilities. Multilaser-excited fluorescence emitted from individual cells is measured at several wavelength ranges, rapidly (up to 5000 cells/min), with high sensitivity and accuracy. The following applications of LSC are reviewed: (1) identification of cells that differ in degree of chromatin condensation (e.g., mitotic or apoptotic cells or lymphocytes vs granulocytes vs monocytes); (2) detection of translocation between cytoplasm vs nucleus or nucleoplasm vs nucleolus of regulatory molecules such as NF-kappaB, p53, or Bax; (3) semiautomatic scoring of micronuclei in mutagenicity assays; (4) analysis of fluorescence in situ hybridization; (5) enumeration and morphometry of nucleoli; (6) analysis of phenotype of progeny of individual cells in clonogenicity assay; (7) cell immunophenotyping; (8) visual examination, imaging, or sequential analysis of the cells measured earlier upon their relocation, using different probes; (9) in situ enzyme kinetics and other time-resolved processes; (10) analysis of tissue section architecture; (11) application for hypocellular samples (needle aspirate, spinal fluid, etc.); (12) other clinical applications. Advantages and limitations of LSC are discussed and compared with flow cytometry.

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“…The first assays were established to investigate cultured cells and peripheral blood leukocytes, whereas today the spectrum of applications expanded to formalin‐fixed, paraffin‐embedded material, freshly resected tissue, and fresh frozen tissue (5–9). In general, the preparation of single cells or nuclei from solid tissue blocks was performed with different methods, such as mechanical dissociation, grating cells from a tissue block, touch preparation, touch smears, or fine‐needle aspirate biopsies (10–14). Although these methods are well established for tumor material, they are not useful for brain tissue.…”

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confidence: 99%

Laser scanning cytometry in human brain slices

et al. 2006

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Background: The Laser Scanning Cytometry (LSC) offers quantitative fluorescence analysis of cell suspensions and tissue sections. Methods: We adapted this technique to immunohistochemical labelled human brain slices. Results: We were able to identify neurons according to their labelling and to display morphological structures such as the lamination of the entorhinal cortex. Further, we were able to distinguish between neurons with and without cyclin B1 expression and we could assign the expression of cyclin B1 to the cell islands of layer II and the pyramidal neurons of layer V of the entorhinal cortex

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Determination of ploidy and steroid receptor status in breast cancer by laser scanning cytometry

Cited by 9 publications

References 21 publications

Determination of features indicating progression in atypical squamous cells with undetermined significance

Determination of features indicating progression in atypical squamous cells with undetermined significance

Laser Scanning Cytometry

Laser scanning cytometry in human brain slices

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