The Feulgen reaction 75 years on

Chieco, Pasquale; Derenzini, Massimo

doi:10.1007/s004180050367

Cited by 102 publications

(60 citation statements)

References 67 publications

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“…Thus, with appropriate enlargement, each nucleus is divided into hundreds of independent, single photometric measurements, which are summarized to give the final integrated optical density. 36 Video-image cytometers are highly efficient, very easy to use, and cost less than one-third of the cost of the previously used instruments. This lead to a wide expansion of instrument development for research and clinical applications.…”

Section: Instrumentation and Softwarementioning

confidence: 99%

“…Appropriate tests and performance standards have been published to assure sufficient densitometric linearity. 36 Laser scanning cytometry is a new method of image analysis that was developed during the late 1980s and became available commercially in the early 1990s. 48 The method applied to the analysis of populations of more than 100,000 cells per smear is capable of detecting rare DNA events.…”

Section: Instrumentation and Softwarementioning

confidence: 99%

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Diagnostic and prognostic use of DNA image cytometry in cervical squamous intraepithelial lesions and invasive carcinoma

Böcking

Nguyen

2003

Cancer

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In the fight against cervical malignancy and its precursors, several adjuvant diag- In patients with borderline lesions (mild and moderate dysplasias) of the uterine cervix, cytomorphology alone often is not sufficient for the early and definite cytologic detection of malignancy. Cervical dysplasia describes squamous cells or tissues that potentially may lead to malignancy but do not exhibit sufficient evidence for a definite assumption. Resulting from weakness of morphologic criteria to identify malignant transformation in epithelial cells early and unequivocally, dysplasias are not a disease entity. The widely accepted assumption is that the higher the grade of dysplasia is, the higher the probability of progression to carcinoma is.1 However, as a result of insufficient morphologic criteria, neither histologic nor cytologic evaluation can predict whether a lesion will progress to cancer in an individual patient.2 Rates of regression and progression of cervical dysplasia (positive and negative predictive values) are quite different from one study to another. 1 Insufficient interobserver reproducibility in diagnostic cytology and histology represents another dilemma in the microscopic diagnosis of precancerous cervical intraepithelial lesions.Because the diagnosis of cervical squamous intraepithelial lesions (SILs) is not only poorly reproducible but also of limited biologic meaning for the individual patient, the number of resulting control procedures usually is high. These range from repeated cytologic smears and biopsies to unnecessary operations (conizations). Missed early diagnoses of cancers may result from cytomorphologic uncertainties. This also results in unnecessary costs and avoidable anxiety for the patients.In the late 1970s, zur Hausen suggested that there may be an association between human papillomavirus (HPV) and cervical carcinoma.3 Large numbers of subsequent epidemiologic, clinicopathologic, and molecular studies have linked the presence of specific types of HPV to the development of anogenital carcinoma and its precursors. A recent study estimated the worldwide HPV prevalence in cervical carcinomas at 99.7%. 4 Today, it is widely accepted that HPVs play a critical role in the pathogenesis of most cervical carcinomas and their precursor lesions. The infection of cervical epithelial cells with HPV itself is necessary but still insufficient for neoplastic progression. Other factors, especially genetic alterations, are needed to enter into the process of neoplastic transformation. 5Several adjuvant diagnostic methods currently are proposed to increase the diagnostic accuracy and reproducibility of cytology and histology. These range from clinical procedures, like colposcopy, to laboratory methods, like assays for the detection of HPV DNA and HPV typing 6 or DNA image cytometry (DNA-ICM). 1,7,8 In the current review, we focus on the technical performance and application of DNA-ICM in diagnosis and prognosis of cervical SILs and invasive cervical carcinoma. Biologic BackgroundChromosomal aneuploi...

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Section: Instrumentation and Softwarementioning

confidence: 99%

Section: Instrumentation and Softwarementioning

confidence: 99%

Diagnostic and prognostic use of DNA image cytometry in cervical squamous intraepithelial lesions and invasive carcinoma

Böcking

Nguyen

2003

Cancer

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“…Feulgen technique, the gold standard for stoichiometry in 2-D, [51][52][53] showed the best results in 3-D as determined by the highest accuracy (least MSE) to FCM and least measurement variance. We optimized the staining time of acidic thionin for stoichiometry and have shown that 30 to 45 min of staining time is the next best stain for stoichiometry.…”

Section: Discussionmentioning

confidence: 99%

Three-dimensional DNA image cytometry by optical projection tomographic microscopy for early cancer diagnosis

Agarwal

Biancardi

Patten³

et al. 2014

J. Med. Imag

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Abstract. Aneuploidy is typically assessed by flow cytometry (FCM) and image cytometry (ICM). We used optical projection tomographic microscopy (OPTM) for assessing cellular DNA content using absorption and fluorescence stains. OPTM combines some of the attributes of both FCM and ICM and generates isometric highresolution three-dimensional (3-D) images of single cells. Although the depth of field of the microscope objective was in the submicron range, it was extended by scanning the objective's focal plane. The extended depth of field image is similar to a projection in a conventional x-ray computed tomography. These projections were later reconstructed using computed tomography methods to form a 3-D image. We also present an automated method for 3-D nuclear segmentation. Nuclei of chicken, trout, and triploid trout erythrocyte were used to calibrate OPTM. Ratios of integrated optical densities extracted from 50 images of each standard were compared to ratios of DNA indices from FCM. A comparison of mean square errors with thionin, hematoxylin, Feulgen, and SYTOX green was done. Feulgen technique was preferred as it showed highest stoichiometry, least variance, and preserved nuclear morphology in 3-D. The addition of this quantitative biomarker could further strengthen existing classifiers and improve early diagnosis of cancer using 3-D microscopy.

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“…Blood samples were collected at 24, 48, 72, and 96 h after dosing. Peripheral blood erythrocytes were evaluated through micronucleus assay as described previously (Chieco & Derenzini, 1999;Hayashi et al, 2000;Schmid, 1975), with minor modifications. Briefly, blood samples were dropped onto slides, airdried overnight, fixed with absolute methanol for 10 min, stained with Schiff's reagent for 60 min, counterstained with Fast Green 1% for 1 min, and mounted with coverslips.…”

Section: Protocol B (Same Dose Evaluated At Different Time Periods)mentioning

confidence: 99%

In vitroandin vivogenotoxic evaluation ofBothrops moojenisnake venom

Zobiole

Caon

Bertól

et al. 2014

Pharmaceutical Biology

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Context: Bothrops moojeni Hoge (Viperidae) venom is a complex mixture of compounds with therapeutic potential that has been included in the research and development of new drugs. Along with the biological activity, the pharmaceutical applicability of this venom depends on its toxicological profile. Objective: This study evaluates the cytotoxicity and genotoxicity of the Bothrops moojeni venom (BMV). Material and methods: The in vitro cytotoxicity and genotoxicity of a pooled sample of BMV was assessed by the MTT and Comet assay, respectively. Genotoxicity was also evaluated in vivo through the micronucleus assay. Results: BMV displayed a 50% cytotoxic concentration (CC 50 ) on Vero cells of 4.09 mg/mL. Vero cells treated with 4 mg/mL for 90 min and 6 h presented significant (p50.05, ANOVA/NewmanKeuls test) higher DNA damage than the negative control in the Comet assay. The lower DNA damage found after 6 h compared with the 90 min treatment suggests a DNA repair effect. Mice intraperitoneally treated with BMV at 10, 30, or 80 mg/animal presented significant genotoxicity (p50.05, ANOVA/Newman-Keuls test) in relation to the negative control after 24 h of treatment. Contrary to the in vitro results, no DNA repair seemed to occur in vivo up to 96 h post-venom inoculation at a dose of 30 mg/animal. Discussion and conclusion: The results show that BMV presents cyto-and genotoxicity depending on the concentration/dose used. These findings emphasize the importance of toxicological studies, including assessment of genotoxicity, in the biological activity research of BMV and/or in the development of BMV-derived products.

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The Feulgen reaction 75 years on

Cited by 102 publications

References 67 publications

Diagnostic and prognostic use of DNA image cytometry in cervical squamous intraepithelial lesions and invasive carcinoma

Diagnostic and prognostic use of DNA image cytometry in cervical squamous intraepithelial lesions and invasive carcinoma

Three-dimensional DNA image cytometry by optical projection tomographic microscopy for early cancer diagnosis

In vitroandin vivogenotoxic evaluation ofBothrops moojenisnake venom

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