Extraction of nuclei from selected regions in paraffin‐embedded tissue

Oud, Peter S.; Hanselaar, Ton; Reubsaet‐Veldhuizen, José A. M.; Meijer, Jos W. R.; Gemmink, Anita H.; Pahlplatz, M. M. M.; Beck, H. L. M.; Vooijs, G. P.

doi:10.1002/cyto.990070615

Cited by 27 publications

(8 citation statements)

References 9 publications

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“…Nuclei extraction was performed from three to four 50-mm sections from representative tumor areas as described by Oud et al (1986). Cytocentrifugation preparations of the nuclei suspension were stained with the Schiff-Feulgen stain, and the DNA content of 200 stained intact nuclei was then measured by use of the CAS 100 System (Cell Analysis, Lombard, IL) (Rondez et al, 1991).…”

Section: Dna Ploidy Analysismentioning

confidence: 99%

Distinct chromosomal aberrations in sinonasal mucosal melanoma as detected by comparative genomic hybridization

Dijk¹,

Sprenger²,

Rombout³

et al. 2002

Genes Chromosomes & Cancer

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Sinonasal mucosal melanomas are the most frequent mucosal melanomas and arise from melanocytes located in the nasal cavity and the paranasal sinuses. The melanoma types, cutaneous melanoma, uveal melanoma, and mucosal melanoma, differ in etiology, geographic distribution, and clinical behavior. Genetic alterations have been previously studied in cutaneous and uveal melanomas but, to the best of our knowledge, not in mucosal melanomas. Comparative genomic hybridization (CGH) was performed on 14 routinely processed sinonasal mucosal melanomas. Furthermore, ploidy analysis was performed on 11 tumors to provide complementary data on the DNA index. The CGH profiles of sinonasal mucosal melanomas show remarkably consistent alterations: chromosome arm 1q is gained in all tumors and gains of 6p and 8q are present in 93 and 57%, respectively. Comparison of CGH data with both the common variants of cutaneous melanoma and uveal melanoma revealed that sinonasal mucosal melanomas harbor a distinct pattern of chromosomal abnormalities. Ploidy analysis also showed that diploid tumors exhibit gains of 1q and alterations of chromosome 6 (3 of 3 cases tested), whereas clear-copy gains and high-copy gains were seen only in triploid and tetraploid tumors (6 of 8 cases tested). This indicates that alteration of chromosomes 1 and 6 may precede polyploidization and formation of clear-copy gains and high-copy gains.

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Section: Dna Ploidy Analysismentioning

confidence: 99%

Distinct chromosomal aberrations in sinonasal mucosal melanoma as detected by comparative genomic hybridization

Dijk¹,

Sprenger²,

Rombout³

et al. 2002

Genes Chromosomes & Cancer

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“…The specimen preparation procedure, which has been described in detail elsewhere, includes extraction of intact nuclei from selected areas of paraffin-embedded tissue (14,21,22). In short, it includes selection of the CIN I11 and INV lesions from 50 pm "thick" paraffinembedded tissue sections, using a dissecting microscope.…”

Section: Cell Materialsmentioning

confidence: 99%

DNA ploidy and cytophotometric analysis of cervical intraepithelial neoplasia grade III and invasive squamous cell carcinoma

1990

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Cervical intraepithelial neoplasia grade I11 (CIN 111) and squamous cell carcinoma (INV) were examined using DNA ploidy and cytophotometric analysis. Based on hysterectomy, exconisation, and biopsy material from 69 patients in two age categories, analysis was performed in nuclei isolated from selected areas of paraffin-embedded tissue. High percentages of DNA-diploidy in INV lesions were found mainly in the group of patients age 45 years or younger. CIN I11 lesions in women age 46 or older demonstrated high percentages of DNAaneuploidy. DNA-polyploidy was most frequent in CIN I11 lesions in the younger age category. The results of cytophotometric analysis indicated that the overall mean values of 16 nuclear photometric features discriminated significantly between the whole groups of CIN I11 (n = 37) and INV (n = 32). On an individual patient level, however, the mean feature values showed a large overlap. Based on the results of a stepwise linear discriminant analysis of patient mean values, a combination of geometrical and runlength texture features was used to discriminate between CIN I11 and INV lesions. The correct classification rate was highest in the category of patients in the older age category. The results of this study indicate age related differences in CIN I11 and invasive squamous cell carcinoma, and they may be of help in assessing cytophotometric features in the study of progressive and non-progressive CIN lesions.Key terms: Age, paraffin-embedded sample, cytospin, progressive potential Cervical intraepithelial neoplasia grades I to I11 (CIN 1-111 = dysplasia and carcinoma in situ) are potentially precancerous lesions. They can progress to invasive squamous cell carcinoma, but can also persist or return to normal. Earlier follow-up studies of light microscopically detected CIN I and I1 lesions have shown varying frequencies of progression, persistence, and regression (8,9,12,13,19,20,26,28). The end point of most follow-up studies of CIN I and I1 was the development of CIN 111 ( = severe dysplasia and carcinoma in situ), at which time the patients were treated or left the study. Data on the follow-up of CIN 111 itself are less frequent but generally indicate that a relatively small number of CIN I11 lesions will eventually progress to a n invasive squamous cell carcinoma (9,12,13,18,19). Light microscopically, CIN I11 can not be subdivided into progressive, persistent, or regressive subtypes, and as a consequence all cases of CIN I11 are treated as essentially premalignant, potentially progressive lesions. An alternative for visual light microscopic evaluation of CIN I11 lesions is the study of nuclear DNA ploidy and cytophotometric analysis of structural nuclear changes. DNA ploidy analysis, using flow cytometric and histometric techniques, has indicated high percentages of DNA-aneuploidy in CIN 111 lesions (10,16). On this basis some authors concluded that DNA-aneuploidy is a marker for progression in CIN lesions (10). In a recent study on 39 patients with CIN 111, with and without synchronous i...

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“…Various methods have been used to isolate individual nuclei from paraffinembedded tissue for use in diagnostic testing, and modifications of some of these have been suitable for use with the FISH method. 20,[24][25][26][27][28] The extraction of individual nuclei from thick (50-mm) paraffin sections 20 has helped to alleviate some of the technical difficulties associated with the use of thin sections. This method requires a large portion of the available diagnostic tissue, however, and may be undesirable if tissue is limited.…”

Section: Discussionmentioning

confidence: 99%

Tumor Cell Nuclei Extraction From Paraffin-Embedded Lymphoid Tissue for Fluorescence In Situ Hybridization

Rowe

Willmore-Payne

Tripp

et al. 2006

Applied Immunohistochemistry &Amp; Molecular Morphology

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In this study the authors evaluated a technique for isolating intact tumor nuclei from paraffin-embedded lymphoma samples before performing FISH testing to detect the lymphoma-specific trans-location t(11;14) that defines mantle cell lymphoma. Well-characterized surgical pathology cases of mantle cell lymphoma were identified from pathology archives. Thin sections were cut from the paraffin-embedded tissue blocks. One section was stained using hematoxylin and eosin and an area composed exclusively of malignant cells was identified and marked on the slide. The corresponding area of the tissue block corresponding to this region underwent needle core biopsy, and the tissue was processed to isolate tumor cell nuclei and deposited onto a glass slide. The paired sample preparations underwent routine FISH testing for detection of the t(11;14)(q13;q32) chromosomal trans-location. DNA probe hybridization quality was compared between the tissue and isolated nuclei. Individual tumor cell nuclei were successfully extracted from each of the tissue blocks. The t(11;14) trans-location was detected by FISH in all of the samples diagnosed as mantle cell lymphoma. The hybridization signals found in the nuclei of extracted tumor cells were bright, planar, and easily identified. Detection of signal was superior to that on whole tissue samples, where signals often overlapped or were truncated. This technique produces intact nuclei for analysis, preserves the tissue block for additional studies, and allows sampling of a specific area of the tissue block. This approach may be particularly useful when the amount of diagnostic tissue is limited.

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Extraction of nuclei from selected regions in paraffin‐embedded tissue

Cited by 27 publications

References 9 publications

Distinct chromosomal aberrations in sinonasal mucosal melanoma as detected by comparative genomic hybridization

Distinct chromosomal aberrations in sinonasal mucosal melanoma as detected by comparative genomic hybridization

DNA ploidy and cytophotometric analysis of cervical intraepithelial neoplasia grade III and invasive squamous cell carcinoma

Tumor Cell Nuclei Extraction From Paraffin-Embedded Lymphoid Tissue for Fluorescence In Situ Hybridization

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