The Effect of Primary Fixation with Standard Postfixation and the Duration of Hydrolysis on Nuclear Features in Image Cytometry

Fležar, Margareta Strojan; Doudkine, Alexei; Us‐Krašovec, Marija

doi:10.1155/1998/893975

Cited by 3 publications

(5 citation statements)

References 11 publications

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“…The IOD and specifically nuclear texture features in the cell suspensions from formalin fixed paraffinembedded tissues in our study reach the plateau on hydrolysis curve earlier than cells from fresh tissues fixed mainly by alcohol fixatives [7,22]. It is known that formaldehyde fixation induces the formation of cross-links between DNA and nucleoproteins, which enhances the access of dye to potential dye-binding sites [14].In contrast, previous studies on the effect of formalin fixation on IOD values, where the IOD plateau in the formalin fixed cells appeared later than in the alcohol fixed cells, analysed the cells obtained from fresh tissue by imprinting, or from cell cultures, where no enzymatic disaggregation was used in the preparation procedure.…”

Section: Discussionmentioning

confidence: 51%

“…In contrast, alcohol fixed tissue is less protected against the harmful effect of hydrolysis and depolymerized DNA fragments are extracted earlier. Consequently, IOD decreases and nuclear texture features deteriorate earlier [7,22].…”

Section: Discussionmentioning

confidence: 99%

“…However, there is no substantial information on the effect of hydrolysis treatment on nuclear texture features in the cell suspensions from formalin fixed paraffinembedded tissues. In our previous study on fresh cytologic material, we have shown that the plateau of hydrolysis curve, which represents the optimal time for hydrolysis treatment, was shorter, if nuclear texture feature values instead of IOD values were plotted on x-axis [7].…”

Section: Introductionmentioning

confidence: 99%

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Hydrolysis Profiles of Formalin Fixed Paraffin‐Embedded Tumors Based on IOD (Integrated Optical Density) and Nuclear Texture Feature Measurements

Fležar

Us‐Krašovec²,

Žganec³

et al. 1999

Analytical Cellular Pathology

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The aim of the study was to determine optimal hydrolysis time for the Feulgen DNA staining of archival formalin fixed paraffin‐embedded surgical samples, prepared as single cell suspensions for image cytometric measurements. The nuclear texture features along with the IOD (integrated optical density) of the tumor nuclei were analysed by an automated high resolution image cytometer as a function of duration of hydrolysis treatment (in 5 N HCl at room temperature). Tissue blocks of breast carcinoma, ovarian serous carcinoma, ovarian serous tumor of borderline malignancy and leiomyosarcoma were included in the study. IOD hydrolysis profiles showed plateau between 30 and 60 min in the breast carcinoma and leiomyosarcoma, and between 40 and 60 min in the ovarian serous carcinoma and ovarian serous tumor of borderline malignancy. Most of the nuclear texture features remained stable after 20 min of hydrolysis treatment. Our results indicate that the optimal hydrolysis time for IOD and for nuclear texture feature measurements, was between 40 and 60 min in the cell preparations from tissue blocks of three epithelial and one soft tissue tumor.

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Section: Discussionmentioning

confidence: 51%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Hydrolysis Profiles of Formalin Fixed Paraffin‐Embedded Tumors Based on IOD (Integrated Optical Density) and Nuclear Texture Feature Measurements

Fležar

Us‐Krašovec²,

Žganec³

et al. 1999

Analytical Cellular Pathology

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“…Direct comparison of nuclear texture features between the primary tumors and metastases was not prudent due to initial differences in primary fixation procedures of the two sample types. 17 …”

Section: Resultsmentioning

confidence: 99%

“…Unfortunately, due to the differences in primary fixation procedures, which influence the nuclear features of the two sample types, as we already proven previously, this was inappropriate in the present study. 17 …”

Section: Discussionmentioning

confidence: 99%

Image cytometric nuclear texture features in inoperable head and neck cancer: a pilot study

Strojan-Fležar¹,

Lavrencak²,

Žganec³

et al. 2011

Radiology and Oncology

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BackgroundImage cytometry can measure numerous nuclear features which could be considered a surrogate end-point marker of molecular genetic changes in a nucleus. The aim of the study was to analyze image cytometric nuclear features in paired samples of primary tumor and neck metastasis in patients with inoperable carcinoma of the head and neck.Materials and methods.Image cytometric analysis of cell suspensions prepared from primary tumor tissue and fine needle aspiration biopsy cell samples of neck metastases from 21 patients treated with concomitant radiochemotherapy was performed. Nuclear features were correlated with clinical characteristics and response to therapy.ResultsManifestation of distant metastases and new primaries was associated (p<0.05) with several chromatin characteristics from primary tumor cells, whereas the origin of index cancer and disease response in the neck was related to those in the cells from metastases. Many nuclear features of primary tumors and metastases correlated with the TNM stage.ConclusionsA specific pattern of correlation between well-established prognostic indicators and nuclear features of samples from primary tumors and those from neck metastases was observed. Image cytometric nuclear features represent a promising candidate marker for recognition of biologically different tumor subgroups.

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Automated identification of diploid reference cells in cervical smears using image analysis

Laak¹,

Siebers²,

Cuijpers³

et al. 2002

Cytometry

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Linear and cyclic analogues of cyclolinopeptide A (CLA) with two dipeptide segments (Val5Pro6 and Pro6Pro7) replaced by their tetrazole derivatives were synthesized by the SPPS technique and cyclized using TBTU (O‐(benzotriazol‐1‐yl)‐1,1,3,3‐tetramethyluronium tetrafluoroborate) reagent. The conformational properties of the c(Leu1Ile2Ile3Leu4Val5Pro6 ψ[CN4]Ala7Phe8Phe9) were investigated by NMR and computational techniques. The overall solution structure of this cyclic peptide resembles that observed for the CLA in the solid state. These studies of cyclic tetrazole CLA analogue confirm that the 1,5‐disubstituted tetrazole ring functions as an effective, well‐tolerated cis‐amide bond mimic in solution. The peptides were examined for their immunosuppressive activity in the humoral response test. For cyclic analogues the immunosuppressive activity, at low doses, is equal in magnitude to the activity presented by cyclosporin A and native CLA. The conformational and biological data seem indicate that the ProProPhePhe moiety and the preservation of the CLA backbone conformation are important for immunosuppressive activity. © 2002 Wiley Periodicals, Inc. Biopolymers 63: 343–357, 2002

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The Effect of Primary Fixation with Standard Postfixation and the Duration of Hydrolysis on Nuclear Features in Image Cytometry

Cited by 3 publications

References 11 publications

Hydrolysis Profiles of Formalin Fixed Paraffin‐Embedded Tumors Based on IOD (Integrated Optical Density) and Nuclear Texture Feature Measurements

Hydrolysis Profiles of Formalin Fixed Paraffin‐Embedded Tumors Based on IOD (Integrated Optical Density) and Nuclear Texture Feature Measurements

Image cytometric nuclear texture features in inoperable head and neck cancer: a pilot study

Automated identification of diploid reference cells in cervical smears using image analysis

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