Flat embeddment of monolayer cells, tissue sections, and other cellular materials attached onto glass substrate

1990 ◽  
Vol 48 (3) ◽  
pp. 766-767
Author(s):  
Jeffrey P. Chang ◽  
Jaang J. Wang
Keyword(s):  
Present Report ◽  
Cellular Materials ◽  
Cover Glass ◽  
Tissue Sections ◽  
Embedding Technique ◽  
Exfoliated Cells ◽  
Black Paper ◽  
Cell Concentrations ◽  
Flat Embedding

Flat embeddment of certain specimens for electron microscopy is necessary for three classes of biological materials: namely monolayer cells, tissue sections of paraffin or plastics, as well as cell concentrations, exfoliated cells, and cell smears. The present report concerns a flat-embedding technique which can be applied to all these three classes of materials and which is a modified and improved version of Chang's original methodology.Preparation of coverglasses and microslides. Chemically cleaned coverglasses, 11 × 22 mm or other sizes, are laid in rows on black paper. Ink-mark one coner for identifying the spray-side of the glass for growing cells. Lightly spray with Teflon monomer (Heddy/Contact Inductries, Paterson, NO 07524, U.S.A.) from a pressurized can. Bake the sprayed glasses at 500°F for 45 min on Cover-Glass Ceramic Racks (A. Thomas Co. Philadelphia), for Teflon to polymerize.Monolayer Cells. After sterilization, the Teflon-treated coverglasses, with cells attached, are treated or fixed in situ in Columbia staining dishes (A. Thomas Co., Philadelphia) for subsequent processing.

Involvement of nucleus in the maturation of dengue-2 virus in C6/36 cells

1990 ◽  
Vol 48 (3) ◽  
pp. 912-913
Author(s):  
Jaang J. Wang ◽  
Cheng C. Chen ◽  
Men F. Shaio ◽  
Chia T. Liu ◽  
Chung S. Lee ◽  
...  
Keyword(s):  
Culture Medium ◽  
Cytopathic Effect ◽  
Fetal Bovine Serum ◽  
Virus Particles ◽  
Electron Microscopies ◽  
Embedding Technique ◽  
Fetal Bovine ◽  
Flat Embedding ◽  
20 Nm ◽  
Labeling Method

The involvement of nucleus in the maturation processes of Dengue-2 virus in a mosquito cell line, C6/36 cells, has been identified by the electron microscopy and immunocytochemistry. The C6/36 cells were obtained from ATCC and maintained in MEM culture medium containing 10% fetal bovine serum at 28°C. The cell suspensions or cells grown on teflon-coated coverslips were infected with Dengue-2 virus (107/ml) for various time periods of 2 hours, 3, 6, 8, and 10 days. The cells were then fixed in buffered 1.5% glutaraldehyde, and washed in acetone before immunolabeled with monoclonal antibody. An indirect immunocytochemical labeling method of avidin-biotin complex (ABC) conjugated with peroxidase or gold particles (20 nm in diameter) and a flat embedding technique were used to localize the virus particles.At early stages of infections (before 3 days), there were no virion particles detected. After 6 days and on of infections, cytopathic effect (CPE) was observed and showed positive immuno-peroxidase reactions under the light and electron microscopies.

Not All Cases of Mammary Paget’s Disease are Cytokeratin-7 Positive: A Challenging Diagnosis!

2021 ◽  
pp. 106689692110029
Author(s):  
Kerschen Anja ◽  
Dano Hélène ◽  
Van Eeckhout Pascal ◽  
Marot Liliane ◽  
Van Bockstal Mieke
Keyword(s):  
Present Report ◽  
Paget’S Disease ◽  
Growth Factor Receptor ◽  
Punch Biopsy ◽  
Paget's Disease ◽  
Cytokeratin 7 ◽  
In Situ Carcinoma ◽  
Epidermal Growth ◽  
Mammary Paget’S Disease

Mammary Paget’s disease accounts for 1% to 3% of all breast tumors and manifests as a chronic eczematous lesion of the areolar skin. It can occur without any underlying neoplasia or can be present in association with an underlying invasive and/or in situ carcinoma of the breast. The present report describes a challenging nipple punch biopsy showing an infiltration of the lower third to two-thirds of the epidermis by large, ovoid, neoplastic cells. The morphology was consistent with mammary Paget's disease, although immunohistochemistry for cytokeratin-7 (CK7) was repeatedly negative. This resulted in an initial misdiagnosis and, subsequently, a delay in the patient's follow-up. Additional immunohistochemistry for GATA binding protein 3 (GATA3) and human epidermal growth factor receptor 2 (HER2), as well as a second opinion of a breast pathologist, resulted in the diagnosis of mammary Paget's disease. The aim of this article is to raise awareness among pathologists and prevent them from misdiagnosing CK7-negative Paget disease of the breast.

On The Optimized Nucleation of Near-Single-Crystal Cvd Diamond Film

1995 ◽  
Vol 416 ◽  
Author(s):  
L. C. Chen ◽  
C. C. Juan ◽  
J. Y. Wu ◽  
K. H. Chen ◽  
J. W. Teng
Keyword(s):  
Single Crystal ◽  
Photoelectron Spectroscopy ◽  
Morphological Evolution ◽  
Present Report ◽  
Ex Situ ◽  
Nucleation Density ◽  
Induced Current ◽  
Current Time ◽  
Single Crystal Diamond

ABSTRACTNear-single-crystal diamond films have been obtained in a number of laboratories recently. The optimization of nucleation density by using a bias-enhanced nucleation (BEN) method is believed to be a critical step. However, the condition of optimized nucleation has never been clearly delineated. In the present report, a novel quantitative technique was established to monitor the nucleation of diamond in-situ. Specifically, the induced current was measured as a function of nucleation time during BEN. The timedependence of induced current was studied under various methane concentrations as well as substrate temperatures. The optimized nucleation condition can be unambiguously determined from the current-time plot. Besides the in-situ current probe, ex-situ x-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) were also used to investigate the chemical and morphological evolution. Characteristic XPS and AFM features of optimized nucleation is discussed.

Porcine Repeat Element DNA: In Situ Detection of Xenotransplanted Cells

1995 ◽  
Vol 4 (2) ◽  
pp. 253-256 ◽  
Author(s):  
Henry F. Oettinger ◽  
Amelie Rodrigue-Way ◽  
Joyce J. Bousquet ◽  
Albert S.B. Edge
Keyword(s):  
Southern Blotting ◽  
Host Tissue ◽  
Repeat Element ◽  
Tissue Analysis ◽  
Specific Staining ◽  
Tetrazolium Chloride ◽  
Tissue Sections ◽  
Blue Tetrazolium Chloride ◽  
In Situ Detection

Using a digoxygenin-labelled DNA probe derived from the porcine repeat element PRE-1, we have developed a protocol for the detection of transplanted porcine islets and hepatocytes against a background of murine host tissue. Analysis of this probe by Southern blotting indicated that PRE-1 hybridizes to pig genomic DNA but not to human or mouse DNA. On tissue sections, hybridizing probe was detected using alkaline phosphatase-conjugated anti-digoxygenin antibody visualized with 5-bromo-4-chloro-3-indolyl-phosphate/4-nitro-blue tetrazolium chloride (BCIP/ NBT) substrate. We have demonstrated sensitive and highly specific staining of porcine nuclei in fixed, paraffin embedded tissue sections, and have applied the technique to detect porcine pancreatic islets and hepatocytes transplanted into murine kidney and spleen. Applications of this technique include detection of transplanted cells or organs across a variety of xenogeneic barriers.

Molecular Hybridization of Mouse Satellite DNA-Complementary RNA in Ultrathin Sections Prepared for Electron Microscopy

1974 ◽  
Vol 14 (2) ◽  
pp. 253-261
Author(s):  
J. JACOB ◽  
KATHERINE GILLIES ◽  
D. MACLEOD ◽  
K. W. JONES
Keyword(s):  
Electron Microscopy ◽  
Satellite Dna ◽  
Similar Distribution ◽  
Tissue Sections ◽  
Satellite Sequences ◽  
Interphase Cells ◽  
Nucleolar Chromatin ◽  
Ultrathin Sections ◽  
Mouse Satellite Dna

The feasibility of in situ hybridization in tissue sections prepared for electron microscopy has been examined using mouse satellite DNA-complementary RNA and mouse L cells. The results obtained are encouraging, although certain technical aspects require further clarification. In interphase cells, hybrid-forming sites occur in chromatin patches positioned along the nuclear envelope. It is also confirmed that satellite DNA occurs in nucleolus-associated chromatin. The results suggest that satellite sequences are present in intranucleolar and peri-nucleolar chromatin. A similar distribution is indicated for ribosomal cistrons.

scholarly journals A new method to detect apoptosis in paraffin sections: in situ end-labeling of fragmented DNA.

1993 ◽  
Vol 41 (1) ◽  
pp. 7-12 ◽  
Author(s):  
J H Wijsman ◽  
R R Jonker ◽  
R Keijzer ◽  
C J van de Velde ◽  
C J Cornelisse ◽  
...  
Keyword(s):  
Cell Death ◽  
Programmed Cell Death ◽  
Staining Method ◽  
Morphological Characteristics ◽  
Image Cytometry ◽  
Apoptotic Cells ◽  
Dna Breaks ◽  
Paraffin Sections ◽  
Tissue Sections

Apoptosis (programmed cell death) can be difficult to detect in routine histological sections. Since extensive DNA fragmentation is an important characteristic of this process, visualization of DNA breaks could greatly facilitate the identification of apoptotic cells. We describe a new staining method for formalin-fixed, paraffin-embedded tissue sections that involves an in situ end-labeling (ISEL) procedure. After protease treatment to permeate the tissue sections, biotinylated nucleotides are in situ incorporated into DNA breaks by polymerase and subsequently stained with DAB via peroxidase-conjugated avidin. Staining of cells with the morphological characteristics of apoptosis was demonstrated in tissues known to exhibit programmed cell death, i.e., prostate and uterus after castration, tumors, lymph node follicles, and embryos. Apoptotic cells could be discriminated morphologically from areas of labeled necrotic cells, in which DNA degradation also occurs. Because apoptosis is relatively easily recognized in H&E-stained sections of involuting prostates of castrated rats, we used this model system to validate the ISEL method for the quantification of apoptotic cells. A high correlation was found between the fractions of ISEL-labeled cells and the fractions of apoptotic cells that were morphologically determined in adjacent sections. We conclude that ISEL is a useful technique for quantification of apoptosis in paraffin sections, especially for those tissues in which morphological determination is difficult. Furthermore, this new staining method enables the use of automated image cytometry for evaluating apoptosis.

scholarly journals AN INTERVENTIONAL STUDY TO ASSESS THE CYTOLOGICAL AND COLPOSCOPIC CHANGES AFTER INTRAUTERINE CONTRACEPTIVE DEVICES INSERTION IN THE DEPARTMENT OF OBSTETRICS GYNAECOLOGY, SMS MEDICAL COLLEGE, JAIPUR

2021 ◽  
Vol 5 (7) ◽  
Author(s):  
Mamta Prajapat ◽  
Suniti Verma ◽  
R N Sehra ◽  
Jyoti Saini
Keyword(s):  
Trichomonas Vaginalis ◽  
Pap Smear ◽  
Medical College ◽  
Contraceptive Devices ◽  
Exfoliated Cells ◽  
Increased Risk ◽  
Intrauterine Contraceptive ◽  
Long Term Follow Up

Background: Cytology examines exfoliated cells, while colposcopy examines the changes that occur in the vascular structure of the cervix5. Patients with an abnormal pap smear result or patients with a suspicious-looking cervix even if they had a negative pap smear, should be evaluated by colposcopy and colposcopy directed biopsy. Methods: Interventional Prospective study conducted at Department of Obstetrics and Gynaecology, SMS Medical College & associated Hospitals, Jaipur. Results: In this study, among 15 IUCD users, 5 (33.33%) had bacterial vaginosis, 2 (13.33%) had trichomonas vaginalis and 1 (6.66%) were found candidal infection, 1 (6.66%) polymicrobial infection. Conclusion: The present study indicates that there is definite changes in the cervical architecture in IUCD users and the change is also dependent on the duration of use. A simple technique like cytology and colposcopy is highly useful in diagnosing these changes and combating them by early interference whenever necessary. The present study does not reveal any increased risk of cervicitis and dysplasia in IUCD users. We further recommend that a long term follow-up is required to rule out the possibility of any increased risk of dysplasia or carcinoma in-situ. Keywords: IUCD, Colposcopy, Dysplasia.

scholarly journals High resolution spatial transcriptome analysis by photo-isolation chemistry

2020 ◽  
Author(s):  
Mizuki Honda ◽  
Shinya Oki ◽  
Akihito Harada ◽  
Kazumitsu Maehara ◽  
Kaori Tanaka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcriptome Analysis ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Transcriptome Profiling ◽  
Tissue Sections ◽  
Complex Functions ◽  
Multicellular Organisms

ABSTRACTIn multicellular organisms, individual cells are characterized by their gene expression profiles and the spatial interactions among cells enable the elaboration of complex functions. Expression profiling in spatially defined regions is crucial to elucidate cell interactions and functions. Here, we established a transcriptome profiling method coupled with photo-isolation chemistry (PIC) that allows the determination of expression profiles specifically from photo-irradiated regions of whole tissues. PIC uses photo-caged oligodeoxynucleotides for in situ reverse transcription. After photo-irradiation of limited areas, gene expression was detected from at least 10 cells in the tissue sections. PIC transcriptome analysis detected genes specifically expressed in small distinct areas of the mouse embryo. Thus, PIC enables transcriptome profiles to be determined from limited regions at a spatial resolution up to the diffraction limit.

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