Discovery of intracellular 2,8 dihydroxyadenine crystals in bovine lymphatic and hepatic cells

1987 ◽  
Vol 45 ◽  
pp. 906-907
Author(s):  
W. E. Rigsby ◽  
D. M. Hinton ◽  
V. J. Hurst ◽  
P. C. McCaskey
Keyword(s):  
Polarized Light ◽  
Paraffin Sections ◽  
Tissue Samples ◽  
Whole Cells ◽  
Tissue Sections ◽  
Hepatic Cells ◽  
Slaughter House ◽  
Mammalian Tissues ◽  
Carbon Coated ◽  
Cellular Components

Crystalline intracellular inclusions are rarely seen in mammalian tissues and are often difficult to positively identify. Lymph node and liver tissue samples were obtained from two cows which had been rejected at the slaughter house due to the abnormal appearance of these organs in the animals. The samples were fixed in formaldehyde and some of the fixed material was embedded in paraffin. Examination of the paraffin sections with polarized light microscopy revealed the presence of numerous crystals in both hepatic and lymph tissue sections. Tissue sections were then deparaffinized in xylene, mounted, carbon coated, and examined in a Phillips 505T SEM equipped with a Tracor Northern X-ray Energy Dispersive Spectroscopy (EDS) system. Crystals were obscured by cellular components and membranes so that EDS spectra were only obtainable from whole cells. Tissue samples which had been fixed but not paraffin-embedded were dehydrated, embedded in Spurrs plastic, and sectioned.

scholarly journals Intensification of the immunocytochemical reaction by staining both sides of tissue sections.

1987 ◽  
Vol 35 (2) ◽  
pp. 257-260 ◽  
Author(s):  
D R Ciocca ◽  
A O Stati ◽  
L C Fasoli
Keyword(s):  
Incubation Time ◽  
Binding Sites ◽  
Primary Antibody ◽  
Paraffin Sections ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Antigenic Sites ◽  
Tissue Area ◽  
Immunocytochemical Reaction ◽  
Complex Methods

We describe how the efficiency of immunostaining may be increased by staining paraffin sections on both sides. This modification exposes more antigenic binding sites per unit tissue area, as shown by the peroxidase-antiperoxidase and the avidin-biotin-peroxidase complex methods. Using antigen-rich tissue samples, the modified procedure made it possible to use more dilute primary antiserum or to reduce the incubation time of the tissue with the primary antibody. Alternatively, in tissue samples with sparse antigenic sites, the procedure made it possible to visualize and document very weak immunoreactivities.

An enhanced cellular organelle visualization procedure when staining for peroxisomes

1995 ◽  
Vol 53 ◽  
pp. 886-887
Author(s):  
J. W. Horn ◽  
B.J. Dovey-Hartman
Keyword(s):  
Tissue Architecture ◽  
Ultrastructural Examination ◽  
Fixed Tissue ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Visual Identification ◽  
Transmission Electron ◽  
Light Staining ◽  
Cellular Components

Reliable visual identification of peroxisomes is important in developmental, clinical, and investigational research. The current technique employed in most laboratories uses a specific electron dense label for the demonstration of peroxisomes by transmission electron microscopy by applying 3,3'- Diaminobenzidine Tetrahydrochloride (DAB) directly to freshly fixed tissue samples to react with endogenous peroxisomal catalase. After routine processing, ultrastructural examination of tissue sections is conducted either with light staining or without post-staining of grids. While peroxisomes are easily identified using this method, remaining tissue architecture is difficult to visualize due to the opacity of the tissue. Additionally, if grids are post-stained with heavy metal solutions, they must be modified to allow for enough staining to visualize cellular components without compromising the quality of the peroxisome label. We will describe a technique whereby DAB-reacted tissues are stained with a postfixative solution including potassium ferricyanide that imparts density to cell membranes and cellular components thereby enhancing identification and interpretion of data.

A Simple Correlative Technique for Morphologic and Energy-Dispersive Analysis of Glass-Mounted Paraffin Sections

1996 ◽  
Vol 54 ◽  
pp. 308-309
Author(s):  
K. W. Baker ◽  
L. King ◽  
R. Walker ◽  
I. Piscopo ◽  
A. Smith
Keyword(s):  
Short Note ◽  
Polarized Light ◽  
Energy Dispersive ◽  
Kidney Cortex ◽  
Paraffin Sections ◽  
Tissue Sections ◽  
Glass Slides ◽  
Empirical Determination ◽  
Unknown Structure

Tissue sections, smears, and many other varied types of specimen are often mounted on glass slides for light microscopic (LM) evaluation and analysis. These same preparations’ using gold/palladiumcoated glass slides as specimen mounts, are also well suited to correlative scanning electron microscopy (SEM) and, energy dispersive X-ray analysis (SEM/EDX). The following short note describes a procedure for glass-mounted specimens that provides slides suitable for combined LM, SEM and SEM/EDX characterization. In addition, the method establishes a specimen-based reference point for the empirical determination of optimum electron probe depth and accelerating voltage for SEM/EDX analysis.For illustrative purposes we used sections of mammalian kidney cortex known to be heavily laden with crystalline deposits of unknown structure and composition.In each sample, birefringent crystalline material was initially observed in hematoxylin and eosinstained paraffin sections using bright field polarized light microscopy (Fig. 1).

In situ microprobe quantitation of inorganic particle burden in paraffin sections of lung tissue

1988 ◽  
Vol 46 ◽  
pp. 990-991
Author(s):  
Jerrold L. Abraham
Keyword(s):  
Lung Tissue ◽  
Quantitative Information ◽  
Particulate Material ◽  
Paraffin Sections ◽  
Tissue Samples ◽  
Qualitative And Quantitative ◽  
The Past ◽  
Quantitative Analyses ◽  
Data Result

Inorganic particulate material of diverse types is present in the ambient and occupational environment, and exposure to such materials is a well recognized cause of some lung disease. To investigate the interaction of inhaled inorganic particulates with the lung it is necessary to obtain quantitative information on the particulate burden of lung tissue in a wide variety of situations. The vast majority of diagnostic and experimental tissue samples (biopsies and autopsies) are fixed with formaldehyde solutions, dehydrated with organic solvents and embedded in paraffin wax. Over the past 16 years, I have attempted to obtain maximal analytical use of such tissue with minimal preparative steps. Unique diagnostic and research data result from both qualitative and quantitative analyses of sections. Most of the data has been related to inhaled inorganic particulates in lungs, but the basic methods are applicable to any tissues. The preparations are primarily designed for SEM use, but they are stable for storage and transport to other laboratories and several other instruments (e.g., for SIMS techniques).

Low temperature x-ray microanalysis of frozen-hydrated tobacco leaves

1984 ◽  
Vol 42 ◽  
pp. 284-285
Author(s):  
S. Edith Taylor ◽  
Patrick Echlin ◽  
May McKoon ◽  
Thomas L. Hayes
Keyword(s):  
Low Temperature ◽  
Lemna Minor ◽  
Root Tips ◽  
Tobacco Leaves ◽  
X Ray ◽  
Whole Cells ◽  
Carbon Coated ◽  
The Right ◽  
Beam Currents ◽  
The University

Low temperature x-ray microanalysis (LTXM) of solid biological materials has been documented for Lemna minor L. root tips. This discussion will be limited to a demonstration of LTXM for measuring relative elemental distributions of P,S,Cl and K species within whole cells of tobacco leaves.Mature Wisconsin-38 tobacco was grown in the greenhouse at the University of California, Berkeley and picked daily from the mid-stalk position (leaf #9). The tissue was excised from the right of the mid rib and rapidly frozen in liquid nitrogen slush. It was then placed into an Amray biochamber and maintained at 103K. Fracture faces of the tissue were prepared and carbon-coated in the biochamber. The prepared sample was transferred from the biochamber to the Amray 1000A SEM equipped with a cold stage to maintain low temperatures at 103K. Analyses were performed using a tungsten source with accelerating voltages of 17.5 to 20 KV and beam currents from 1-2nA.

scholarly journals Rapid brain structure and tumour margin detection on whole frozen tissue sections by fast multiphotometric mid-infrared scanning

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tim Kümmel ◽  
Björn van Marwick ◽  
Miriam Rittel ◽  
Carina Ramallo Guevara ◽  
Felix Wühler ◽  
...  
Keyword(s):  
Imaging System ◽  
Frozen Section ◽  
Computational Effort ◽  
Primary Hepatocellular Carcinoma ◽  
Measuring System ◽  
Sufficient Information ◽  
Tissue Samples ◽  
Mid Infrared ◽  
Tissue Sections ◽  
Frozen Tissue

AbstractFrozen section analysis is a frequently used method for examination of tissue samples, especially for tumour detection. In the majority of cases, the aim is to identify characteristic tissue morphologies or tumour margins. Depending on the type of tissue, a high number of misdiagnoses are associated with this process. In this work, a fast spectroscopic measurement device and workflow was developed that significantly improves the speed of whole frozen tissue section analyses and provides sufficient information to visualize tissue structures and tumour margins, dependent on their lipid and protein molecular vibrations. That optical and non-destructive method is based on selected wavenumbers in the mid-infrared (MIR) range. We present a measuring system that substantially outperforms a commercially available Fourier Transform Infrared (FT-IR) Imaging system, since it enables acquisition of reduced spectral information at a scan field of 1 cm2 in 3 s, with a spatial resolution of 20 µm. This allows fast visualization of segmented structure areas with little computational effort. For the first time, this multiphotometric MIR system is applied to biomedical tissue sections. We are referencing our novel MIR scanner on cryopreserved murine sagittal and coronal brain sections, especially focusing on the hippocampus, and show its usability for rapid identification of primary hepatocellular carcinoma (HCC) in mouse liver.

Visualisation of GABAergic neurons and synapses in the rat brain using immunohistochemistry for two forms of glutamate decarboxylase

2021 ◽  
Vol 21 (2) ◽  
pp. 63-73
Author(s):  
Valeria A. Razenkova ◽  
Dmitrii E. Korzhevskii
Keyword(s):  
Brain Tissue ◽  
Glutamate Decarboxylase ◽  
Brain Structures ◽  
Gabaergic Neurons ◽  
Laboratory Animals ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Background Staining ◽  
The Central Nervous System ◽  
Rat Forebrain

BACKGROUND: Taking into account the importance of GABAergic brain system research and also the opportunity to achieve specific and accurate results in laboratory studies using immunohistochemical approaches, it seems important to have a reliable method of visualization GABA-synthesizing cells, their projections and synapses, for the morphofunctional analysis of GABAergic system both in normal conditions and in the experimental pathology. AIM: The aim of the study was to visualize analyze GABAergic neurons and synapses within rats brain using three different antibody types against glutamate decarboxylase and to identify the optimal conditions for reaction performing. MATERIALS AND METHODS: The study was performed on paraffin brain tissue sections of 5 adult Wistar rats. Immunohistochemical reactions using three antibody types against glutamate decarboxylase isoform 67 (GAD67) and glutamate decarboxylase isoform 65 (GAD65) were performed. Additional controls on C57/Bl6 mice and Chinchilla rabbits brain samples were also carried out. RESULTS: Antibodies used in the research made it possible to achieve high quality of GABAergic structures visualizing without increasing background staining. At the same time different antibody types are distinct in their efficacy to perform immunohistochemistry reaction on laboratory animal brain tissue samples. By performing additional controls, we discovered that there is necessary to adsorb secondary reagents immunoglobulins in order to eliminate nonspecific staining. It was found that GAD67 and GAD65 distribution in rat forebrain structures is different. It was stated that GAD67 immunohistochemistry most completely reveals GABAergic brain structures compared to GAD65 immunhistochemistry. The possibility of determining morphological features of GABAergic neurons and synaptic terminals, as well as performing quantitative analysis, was demonstrated. CONCLUSIONS: The approach proposed makes it possible to specifically visualize GABAergic structures of the central nervous system of different laboratory animals. This could be useful both in fundamental studies and in pathology research.

Rapid Somatic Mutation Testing in Colorectal Cancer by Use of a Fully Automated System and Single-Use Cartridge: A Comparison with Next-Generation Sequencing

2018 ◽  
Vol 3 (2) ◽  
pp. 178-184 ◽  
Author(s):  
M Rabie Al-Turkmani ◽  
Kelley N Godwin ◽  
Jason D Peterson ◽  
Gregory J Tsongalis
Keyword(s):  
Colorectal Cancer ◽  
Next Generation Sequencing ◽  
Ffpe Tissue ◽  
Braf V600e ◽  
Automated System ◽  
Next Generation ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Single Use ◽  
Generation Sequencing

AbstractBackgroundMolecular tests have been increasingly used in the management of various cancers as more targeted therapies are becoming available as treatment options. The Idylla™ system is a fully integrated, cartridge-based platform that provides automated sample processing (deparaffinization, tissue digestion, and DNA extraction) and real-time PCR-based mutation detection with all reagents included in a single-use cartridge. This retrospective study aimed at evaluating both the Idylla KRAS and NRAS-BRAF-EGFR492 Mutation Assay cartridges (research use only) against next-generation sequencing (NGS) by using colorectal cancer (CRC) tissue samples.MethodsForty-four archived formalin-fixed paraffin-embedded (FFPE) CRC tissue samples previously analyzed by targeted NGS were tested on the Idylla system. Among these samples, 17 had a mutation in KRAS proto-oncogene, GTPase (KRAS), 5 in NRAS proto-oncogene, GTPase (NRAS), and 12 in B-Raf proto-oncogene, serine/threonine kinase (BRAF) as determined using the Ion AmpliSeq 50-gene Cancer Hotspot Panel v2. The remaining 10 samples were wild-type for KRAS, NRAS, and BRAF. Two 10-μm FFPE tissue sections were used for each Idylla run, 1 for the KRAS cartridge, and 1 for the NRAS-BRAF-EGFR492 cartridge. All cases met the Idylla minimum tumor content requirement for KRAS, NRAS, and BRAF (≥10%). Assay reproducibility was evaluated by testing commercial controls derived from human cell lines, which had an allelic frequency of 50% and were run in triplicate.ResultsThe Idylla system successfully detected all mutations previously identified by NGS in KRAS (G12C, G12D, G12V, G13D, Q61K, Q61R, A146T), NRAS (G12V, G13R, Q61H), and BRAF (V600E). Compared with NGS, Idylla had a sensitivity of 100%. Analysis of the mutated commercial controls demonstrated agreement with the expected result for all samples and 100% reproducibility. The Idylla system produced results quickly with a turnaround time of approximately 2 h.ConclusionThe Idylla system offers reliable and sensitive testing of clinically actionable mutations in KRAS, NRAS, and BRAF directly from FFPE tissue sections.

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 First evidence of nocardiosis in farmed red drum (Sciaenops ocellatus, Linnaeus) caused by Nocardia seriolae in the Gulf of Mexico

2021 ◽  
Author(s):  
Rodolfo Enrique del Rio-Rodriguez ◽  
Jose Gustavo Ramirez-Paredes ◽  
Sonia Araceli Soto-Rodriguez ◽  
Yechiam Shapira ◽  
Mariana del Jesus Huchin-Cortes ◽  
...  
Keyword(s):  
Current Knowledge ◽  
Red Drum ◽  
Sciaenops Ocellatus ◽  
Internal Organs ◽  
New Host ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Long Rod ◽  
Nocardia Seriolae

ABSTRACTBetween August and December 2013, the offshore cages of a commercial marine farm culturing red drum Sciaenops ocellatus in Campeche Bay were affected by an outbreak of an ulcerative granulomatous disease, reaching 70% cumulative mortality. Thirty-one adults displaying open ulcers on the skin were submitted to our laboratory for diagnosis. Multiple white-yellowish nodules (0.1-0.5 cm in diameter) were present in all internal organs, where the kidney and the spleen were the most severely affected. Histopathological observations of these organs evinced typical granulomatous formations. Gram and Ziehl-Neelsen stains on tissue imprints, bacterial swabs and tissue sections revealed Gram-positive, acid-fast, branching beaded long rod filamentous bacteria. Tissue samples resulted positive for nocardiosis when a Nocardia genus specific nested PCR was used. Definite identification at the species level and taxonomic positioning of the fastidious pathogen was achieved through a specific Nocardia seriolae PCR and by sequencing the gyrB gene of pure isolates. After application of antibiotics during fry production, a posterior follow-up monitoring (from 2014 to 2017) detected mild but recurrent outbreaks of the bacteria with no seasonality pattern. To the extent of our current knowledge, this is the first report of piscine nocardiosis (in a new host -red drum) in Mexico.

Sign in / Sign up

Export Citation Format

Share Document