Search results for "Immunofluorescence"
showing 10 items of 112 documents
Identification of Merkel cells in human skin by specific cytokeratin antibodies:
1984
Merkel cells are special neurosecretory cells which, in adult human skin, are usually very scarce. By immunofluorescence microscopy using antibodies to human cytokeratin polypeptide no. 18, we localized distinct non-keratinocyte cells in the glandular ridges of human fetal and adult plantar epidermis. Using electron and immunofluorescence microscopy, these cells were identified as Merkel cells containing typical neurosecretory granules as well as bundles of intermediate-sized filaments and desmosomes. Two-dimensional gel electrophoresis of the cytoskeletal fractions of microdissected epidermal preparations highly enriched in Merkel cells indicated the presence of cytokeratin polypeptides no…
Monoclonal antibodies to various acidic (type I) cytokeratins of stratified epithelia
1986
Abstract We determined the reactivity of two monoclonal antibodies to cytokeratins that are typically expressed in certain stratified epithelia and several human squamous cell carcinomas using immunoblotting techniques and immunofluorescence microscopy. Antibody Ks 8.12 reacted specifically with cytokeratin polypeptides nos. 13 and 16, and stained noncornified squamous epithelia in a rather uniform way. The examination of diverse human carcinomas showed all squamous cell carcinomas to be positively stained with this antibody, whereas all adenocarcinomas were negative. Another antibody, KK 8.60, reacted with polypeptides nos. 10 and 11, and uniformly stained the suprabasal layers of the epid…
Membrane vesicles shed into the extracellular medium by human breast carcinoma cells carry tumor-associated surface antigens.
1995
We have compared the pattern of surface antigen expression, as detected by monoclonal antibodies (mAbs), in plasma membranes vs shed membrane vesicles of two human breast carcinoma cell lines, MCF-7 and 8701-BC. Antigen expression was detected on cells by immunofluorescence (IF) analysis, whilst, due to their small dimensions, the same technique was not applicable to vesicles. For these structures dot-blot analysis and immunoelectron microscopy (IEM) were employed. When applicable, both cell membranes and membrane vesicles were immunoprecipitated and the precipitate (IP) was analyzed by SDS-PAGE. Cells of both lines expressed HLA class I antigens, epithelial cytokeratins, β1 integrins, CEA …
Comparison of viability assays for Cryptosporidium parvum oocysts after disinfection.
2003
Abstract In order to test various viability assays for Cryptosporidium parvum oocysts were used to infect HCT-8 cells in vitro or baby mice. Infected cells were either stained with fluorescent anti- Cryptosporidium -antibody or lysed and subjected to C. parvum- specific PCR after 48 h. Titrations with infective oocysts were performed and compared to oocysts disinfected with Neopredisan © for 2 h at varying concentrations. Caecal smears and histological sections from infected animals were examined in parallel. The number of foci of parasite development in vitro after immunofluorescent staining correlated well with the infection dose. PCR was less quantifiable and the results were not always …
Changes in tubulin protein expression accompany reorganization of microtubular arrays during cell shaping in barley leaves
1998
Barley (Hordeum vulgare L.) leaves grow from the base and thus exhibit a smooth developmental gradient. Developing mesophyll cells acquire their typical lobed shape synchronously along this gradient. Successive changes in the patterns of cortical microtubules are involved in the shaping process. The changes include formation and dispersal of band-like structures, the establishment of a random network and a dramatic loss of microtubules after completion of cell shaping. When the relative tubulin contents were determined in consecutive segments taken along the leaf, two tubulin maxima were found. They coincided with the establishment of the microtubular bands and the random network, respectiv…
Clastogenic and aneuploidizing effects of antiblastic busulphan revealed by kinetochore immunofluorescence in CHO cells.
1991
We utilized, in CHO cells, the cytoplasm preservation technique to evaluate the micronucleus frequency at different busulphan concentrations, and the indirect immunofluorescence technique, using sera obtained from patients with scleroderma (CREST variant), to analyze if busulphan-induced micronuclei have kinetochores. Results show that this alkylating agent is capable of causing a significant increase of micronuclei in vitro, a great part (40%) of them having CREST-positive kinetochores. These findings confirm the clastogenic effect of busulphan and reveal a considerable capability of this agent to induce aneuploidy. These results are examined taking into account the high incidence of secon…
Treatment with the anti-tumor drugs, cis-platin and mafosfamide, does not affect the structure of prekinetochores in a human breast cancer cell line.…
1996
Abstract The goal of the present article was to determine whether a nuclear parameter, centromere structure of interphase cells, could serve as an indicator to assess cellular damage caused by anti-tumor drugs. These were cis-platin and mafosfamide, which are widely used for the management of solid tumors. To visualize the centromeres, we probed treated and untreated cells of a human breast cancer cell line, MX-1, with a human anti-centromere serum. The serum was obtained from a scleroderma patient and detects antigens associated with prekinetochores of the decondensed chromosomes. The DNA was simultaneously displayed by a specific fluorescent dye. The cells were grown on coverslips, incuba…
Forever Young: Structural Stability of Telomeric Guanine Quadruplexes in the Presence of Oxidative DNA Lesions**
2021
International audience; Human telomeric DNA, in G-quadruplex (G4) conformation, is characterized by a remarkable structural stability that confers it the capacity to resist to oxidative stress producing one or even clustered 8-oxoguanine (8oxoG) lesions. We present a combined experimental/computational investigation, by using circular dichroism in aqueous solutions, cellular immunofluorescence assays and molecular dynamics simulations, that identifies the crucial role of the stability of G4s to oxidative lesions, related also to their biological role as inhibitors of telomerase, an enzyme overexpressed in most cancers associated to oxidative stress.
Deep CNN for IIF Images Classification in Autoimmune Diagnostics
2019
The diagnosis and monitoring of autoimmune diseases are very important problem in medicine. The most used test for this purpose is the antinuclear antibody (ANA) test. An indirect immunofluorescence (IIF) test performed by Human Epithelial type 2 (HEp-2) cells as substrate antigen is the most common methods to determine ANA. In this paper we present an automatic HEp-2 specimen system based on a convolutional neural network method able to classify IIF images. The system consists of a module for features extraction based on a pre-trained AlexNet network and a classification phase for the cell-pattern association using six support vector machines and a k-nearest neighbors classifier. The class…
Deep Convolutional Neural Network for HEp-2 fluorescence intensity classification
2019
Indirect ImmunoFluorescence (IIF) assays are recommended as the gold standard method for detection of antinuclear antibodies (ANAs), which are of considerable importance in the diagnosis of autoimmune diseases. Fluorescence intensity analysis is very often complex, and depending on the capabilities of the operator, the association with incorrect classes is statistically easy. In this paper, we present a Convolutional Neural Network (CNN) system to classify positive/negative fluorescence intensity of HEp-2 IIF images, which is important for autoimmune diseases diagnosis. The method uses the best known pre-trained CNNs to extract features and a support vector machine (SVM) classifier for the …