Fluoronanogold: An Efficient Labeling Reagent for Immunocytochemistry

1998 ◽  
Vol 4 (S2) ◽  
pp. 990-991
Author(s):  
J.M. Robinson
Keyword(s):  
Sample Preparation ◽  
Colloidal Gold ◽  
Detection System ◽  
Large Body ◽  
Ultrastructural Level ◽  
Detection Systems ◽  
Wide Acceptance ◽  
History Of ◽  
A Cell ◽  
High Degree

The field of immunocytochemistry comprises a large body of methodologies whose purpose is to obtain spatial and temporal information about biological samples with a high degree of chemical specificity. The major differences in immunocytochemical methods are: (1) type of sample preparation (i.e., pre-embedding or post-embedding) and (2) type of detection system (i.e., chromogenic, fluorochromes, or particulate). The history of advances in immunocytochemistry can be understood, to a large extent, through the development of the various detection systems. All of these labeling systems have their appropriate applications and are in use today.The introduction of colloidal gold as a particulate immunoprobe revolutionized immunocytochemistry, particularly at the ultrastructural level. The fact that these probes are discrete particles that are electron dense contributes to their wide acceptance. Due to the particulate nature of these probes, labeling density over a given structure or region of a cell can be determined by simple counting procedures.

scholarly journals Ultrathin Cryosections: An Important Tool for Immunofluorescence and Correlative Microscopy1

2003 ◽  
Vol 51 (6) ◽  
pp. 707-714 ◽  
Author(s):  
Toshihiro Takizawa ◽  
John M. Robinson
Keyword(s):  
Electron Microscopy ◽  
Colloidal Gold ◽  
Ultrathin Section ◽  
Detection System ◽  
Placental Tissue ◽  
Counting Procedure ◽  
Fluorescence And Electron Microscopy ◽  
Ultrathin Cryosections ◽  
High Degree ◽  
Number Of Particles

Here we show that ultrathin cryosections of placental tissue can be used as a substrate in immunofluorescence experiments. A high degree of spatial resolution can be achieved in these preparations because there is essentially no out-of-focus fluorescence. Therefore, immunofluorescence microscopy using ultrathin cryosections provides a very useful method for determining the precise subcellular localization of antigens in tissues. In addition, ultrathin cryosections of placenta also serve as a substrate for correlative immunofluorescence and immunoelectron microscopy using FluoroNanogold as the detection system. In correlative microscopy, the exact same structures in the same ultrathin section were observed by both fluorescence and electron microscopy. Using a particle counting procedure and electron microscopy, we compared the labeling obtained with colloidal gold and FluoroNanogold and found a higher number of particles with silver-enhanced FluoroNanogold than with colloidal gold.

Ultrastructure of melanocyte-keratinocyte interactions and pigment transfer in vitro

1978 ◽  
Vol 36 (2) ◽  
pp. 650-651
Author(s):  
Raul I. Garcia ◽  
Evelyn A. Flynn ◽  
George Szabo
Keyword(s):  
Direct Injection ◽  
Skin Pigmentation ◽  
Freeze Fracture ◽  
Pigment Granule ◽  
Time Lapse ◽  
Ultrastructural Level ◽  
Lanthanum Tracer ◽  
A Cell

Skin pigmentation in mammals involves the interaction of epidermal melanocytes and keratinocytes in the structural and functional unit known as the Epidermal Melanin Unit. Melanocytes(M) synthesize melanin within specialized membrane-bound organelles, the melanosome or pigment granule. These are subsequently transferred by way of M dendrites to keratinocytes(K) by a mechanism still to be clearly defined. Three different, though not necessarily mutually exclusive, mechanisms of melanosome transfer have been proposed: cytophagocytosis by K of M dendrite tips containing melanosomes, direct injection of melanosomes into the K cytoplasm through a cell-to-cell pore or communicating channel formed by localized fusion of M and K cell membranes, release of melanosomes into the extracellular space(ECS) by exocytosis followed by K uptake using conventional phagocytosis. Variability in methods of transfer has been noted both in vivo and in vitro and there is evidence in support of each transfer mechanism. We Have previously studied M-K interactions in vitro using time-lapse cinemicrography and in vivo at the ultrastructural level using lanthanum tracer and freeze-fracture.

Ultrastructural Localization of Antigens by Capillary Action Immunocytochemistry

1996 ◽  
Vol 54 ◽  
pp. 40-41
Author(s):  
László G. Kömüves
Keyword(s):  
San Francisco ◽  
Colloidal Gold ◽  
Specific Binding ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Adhesive Tape ◽  
Distilled Water ◽  
Capillary Action ◽  
Rabbit Igg

Light microscopic immunohistochemistry based on the principle of capillary action staining is a widely used method to localize antigens. Capillary action immunostaining, however, has not been tested or applied to detect antigens at the ultrastructural level. The aim of this work was to establish a capillary action staining method for localization of intracellular antigens, using colloidal gold probes.Post-embedding capillary action immunocytochemistry was used to detect maternal IgG in the small intestine of newborn suckling piglets. Pieces of the jejunum of newborn piglets suckled for 12 h were fixed and embedded into LR White resin. Sections on nickel grids were secured on a capillary action glass slide (100 μm wide capillary gap, Bio-Tek Solutions, Santa Barbara CA, distributed by CMS, Houston, TX) by double sided adhesive tape. Immunolabeling was performed by applying reagents over the grids using capillary action and removing reagents by blotting on filter paper. Reagents for capillary action staining were from Biomeda (Foster City, CA). The following steps were performed: 1) wet the surface of the sections with automation buffer twice, 5 min each; 2) block non-specific binding sites with tissue conditioner, 10 min; 3) apply first antibody (affinity-purified rabbit anti-porcine IgG, Sigma Chem. Co., St. Louis, MO), diluted in probe diluent, 1 hour; 4) wash with automation buffer three times, 5 min each; 5) apply gold probe (goat anti-rabbit IgG conjugated to 10 nm colloidal gold, Zymed Laboratories, South San Francisco, CA) diluted in probe diluent, 30 min; 6) wash with automation buffer three times, 5 min each; 7) post-fix with 5% glutaraldehyde in PBS for 10 min; 8) wash with PBS twice, 5 min each; 9) contrast with 1% OSO4 in PBS for 15 min; 10) wash with PBS followed by distilled water for5 min each; 11) stain with 2% uranyl acetate for 10 min; 12) stain with lead citrate for 2 min; 13) wash with distilled water three times, 1 min each. The glass slides were separated, and the grids were air-dried, then removed from the adhesive tape. The following controls were used to ensure the specificity of labeling: i) omission of the first antibody; ii) normal rabbit IgG in lieu of first antibody; iii) rabbit anti-porcine IgG absorbed with porcine IgG.

MINING THE ARCHIVES: A POTTERY SEQUENCE FOR BORNO, NIGERIA

2003 ◽  
Vol 1 (1) ◽  
pp. 39-76 ◽  
Author(s):  
Graham Connah ◽  
S.G.H. Daniels
Keyword(s):  
Large Body ◽  
Archaeological Research ◽  
Independent Data ◽  
Frankfurt Am Main ◽  
Archaeological Pottery ◽  
New Findings ◽  
History Of ◽  
1960S And 1970S ◽  
The 1960S ◽  
Johann Wolfgang Goethe

New archaeological research in Borno by the Johann-Wolfgang-Goethe-Universität, Frankfurt am Main, Germany, has included the analysis of pottery excavated from several sites during the 1990s. This important investigation made us search through our old files for a statistical analysis of pottery from the same region, which although completed in 1981 was never published. The material came from approximately one hundred surface collections and seven excavated sites, spread over a wide area, and resulted from fieldwork in the 1960s and 1970s. Although old, the analysis remains relevant because it provides a broad geographical context for the more recent work, as well as a large body of independent data with which the new findings can be compared. It also indicates variations in both time and space that have implications for the human history of the area, hinting at the ongoing potential of broadscale pottery analysis in this part of West Africa and having wider implications of relevance to the study of archaeological pottery elsewhere.

THE R.J. CHAMBERS COLLECTION: AN “ARCHIVIST'S” REVELATIONS OF 20th CENTURY ACCOUNTING THOUGHT AND PRACTICE

2006 ◽  
Vol 33 (1) ◽  
pp. 145-166 ◽  
Author(s):  
Graeme W. Dean ◽  
Peter W. Wolnizer ◽  
Frank L. Clarke
Keyword(s):  
Prima Facie ◽  
Accounting Education ◽  
Hard Copy ◽  
Accounting History ◽  
Research Areas ◽  
Archival Source ◽  
Inflation Accounting ◽  
History Of ◽  
High Degree ◽  
Accounting Standard Setting

A major, unique accounting archival source, the R.J. Chambers Collection comprises both hard copy and, utilizing cutting-edge search technology, internet accessible materials. From his academic beginnings, Chambers was an orderly person, an archivist of the extensive and varied evidence that underpinned his proposals for accounting reform. Opening research areas for accounting biography, the development of accounting thought, the history of accounting institutions, prosopography, public sector accounting history, and comparative international accounting history are foremost amongst the myriad justifications for seeking to unravel the accounting history “lodes” in archives such as the Goldberg, Chambers, and Briloff Collections [Potter, 2003]. The archiving of the meticulously kept Chambers papers from 1947–1999 provides an opportunity for unfolding the background to events previously withheld from accounting history scholars. Professional episodes in relation to inflation accounting, standard setting, proposals to reform accounting education, and the like that appeared prima facie to be worth investigating are now open to scrutiny from a different angle, with a different type of evidence available in this Collection. This Collection provides a high degree of archival provenance. In particular, it represents an orderly retention of past documentation of what Chambers wrote, and perhaps uniquely for accounting historians, received; thus, providing an extensive window from which to examine the disorderly present environment of acounting.

scholarly journals Euglycemic Diabetic Ketoacidosis after a Single Dose of Empagliflozin in a Patient with Pancreatitis

2021 ◽  
Vol 11 (2) ◽  
pp. 216-218
Author(s):  
Marta Brandão Calçada ◽  
Luís Fernandes ◽  
Rita Soares Costa ◽  
Sara Montezinho ◽  
Filipa Martins Duarte ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Diabetic Ketoacidosis ◽  
Drug Class ◽  
Clinical Suspicion ◽  
Laboratory Findings ◽  
Sodium Glucose Cotransporter ◽  
History Of ◽  
High Degree ◽  
Euglycemic Diabetic Ketoacidosis

Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are the most recently approved drug class for the treatment of type 2 diabetes mellitus (T2D). Although they are largely well-tolerated, their intake has been associated with euglycemic diabetic ketoacidosis (DKA) in some rare cases. We report the case of a 70-year-old male with type 2 diabetes and no history of DKA, who started therapy with empagliflozin one day before presenting with acute pancreatitis and laboratory findings consistent with euglycemic DKA. SGLT2i can induce euglycemic DKA from the first dose. Given the atypical presentation, a high degree of clinical suspicion is required to recognize this complication.

929 Not Another Abscess - A Case of Streptococcal Myositis Misdiagnosed as a Right Axillary Abscess

2021 ◽  
Vol 108 (Supplement_2) ◽  
Author(s):  
O Oyende ◽  
J Jackman
Keyword(s):  
White Cell Count ◽  
General Surgeon ◽  
High Dose ◽  
General Anaesthetic ◽  
Microbiological Analysis ◽  
Reactive Protein ◽  
Affected Tissue ◽  
Life Threatening ◽  
History Of ◽  
High Degree

Abstract Introduction Streptococcal myositis is a rare form of infectious myositis caused by Lansfield A beta-haemolytic streptococci. It is characterised by rapidly spreading inflammation that can result in severe systemic toxicity and necrosis of the affected tissue if not diagnosed and aggressively treated. Presentation We report a case of a 42-year-old male who presented with a one-week history of worsening right axillary swelling that progressed to painful swelling of his arm. Inflammatory markers were significantly elevated with a white cell count of 17 ×109/L and C-reactive protein of 212 mg/L. On examination, a fluctuant axillary swelling was appreciated, and a decision was made for incision and drainage under general anaesthetic. Intraoperative aspiration of his arm revealed copious purulent fluid prompting intraoperative orthopaedic consult and exploration of the anterior compartment in which there was extensive involvement of the biceps muscle. The microbiological analysis revealed gram-positive cocci in chains, and microbiology advice sought for tailoring of antibiotic regimen. He has recovered well. Discussion Though uncommon, the emergency general surgeon should have a high degree of suspicion when evaluating soft tissue infections to avert potentially disastrous outcomes. Conclusion Early diagnosis, aggressive management with high-dose intravenous antibiotics, and surgical debridement are principles to treat this rare, life-threatening infection.

scholarly journals The Return of National Self-Sufficiency? Excavating Autarkic Thought in a De-Globalizing Era

2021 ◽  
Author(s):  
Eric Helleiner
Keyword(s):  
Intellectual History ◽  
International Political Economy ◽  
Economic Thought ◽  
John Maynard Keynes ◽  
New Politics ◽  
Self Sufficiency ◽  
National Economic ◽  
History Of ◽  
High Degree ◽  
Rich History

Abstract As the global crisis triggered by the COVID-19 virus unfolded, The Economist magazine published a cover in May 2020 titled “Goodbye globalization: the dangerous lure of self-sufficiency.” The title summed up well the new political interest in the ideology of national economic self-sufficiency in the pandemic context. Unfortunately, contemporary textbooks in the field of international political economy (IPE) say little about this kind of “autarkic” thought. No survey of the history of autarkic thought exists even within specialist IPE literature or in the fields of intellectual history and the history of economic thought. Filling this gap in existing scholarship, this article highlights a rich history of autarkic thought that includes the ideas of famous thinkers such as Jean-Jacques Rousseau, Johann Fichte, Mohandas Gandhi, and John Maynard Keynes. Three core rationales for a high degree of national self-sufficiency have been advanced in the past: (1) insulation from foreign economic influence, (2) insulation from foreign political and/or cultural influence, and (3) the promotion of international peace. At the same time, considerable disagreements have existed among autarkists about some of these rationales and their relative importance, as well as about the precise meaning of national self-sufficiency. These disagreements stemmed not just from differences in their specific goals but also from the different conditions across time and space in which autarkic thought was developed. In addition to improving understanding of the autarkic ideological tradition, this article contributes to emerging scholarship attempting to overcome Western-centrism in IPE scholarship as well as literature exploring the new politics of de-globalization in the current era.

scholarly journals Multi-Locus Selection and the Structure of Variation at the white Gene of Drosophila melanogaster

Genetics ◽  
1996 ◽  
Vol 144 (2) ◽  
pp. 635-645 ◽  
Author(s):  
David A Kirby ◽  
Wolfgang Stephan
Keyword(s):  
Drosophila Melanogaster ◽  
White Gene ◽  
Transcriptional Unit ◽  
North American Population ◽  
Evolutionary Mechanisms ◽  
Locus Selection ◽  
Neutral Equilibrium ◽  
History Of ◽  
High Degree ◽  
Very High

Abstract We surveyed sequence variation and divergence for the entire 5972-bp transcriptional unit of the white gene in 15 lines of Drosophila melanogaster and one line of D. simulans. We found a very high degree of haplotypic structuring for the polymorphisms in the 3′ half of the gene, as opposed to the polymorphisms in the 5′ half. To determine the evolutionary mechanisms responsible for this pattern, we sequenced a 1612-bp segment of the white gene from an additional 33 lines of D. melanogaster from a European and a North American population. This 1612-bp segment encompasses an 834bp region of the white gene in which the polymorphisms form high frequency haplotypes that cannot be explained by a neutral equilibrium model of molecular evolution. The small number of recombinants in the 834bp region suggests epistatic selection as the cause of the haplotypic structuring, while an investigation of nucleotide diversity supports a directional selection hypothesis. A multi-locus selection model that combines features from both-hypotheses and takes the recent history of D. melanogaster into account may be the best explanation for these data.

scholarly journals MEDALT: single-cell copy number lineage tracing enabling gene discovery

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Fang Wang ◽  
Qihan Wang ◽  
Vakul Mohanty ◽  
Shaoheng Liang ◽  
Jinzhuang Dou ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Single Cell ◽  
Copy Number ◽  
Speciation Analysis ◽  
Population Based ◽  
Lineage Tracing ◽  
Breast Cancer Patients ◽  
Lineage Tree ◽  
History Of ◽  
A Cell

AbstractWe present a Minimal Event Distance Aneuploidy Lineage Tree (MEDALT) algorithm that infers the evolution history of a cell population based on single-cell copy number (SCCN) profiles, and a statistical routine named lineage speciation analysis (LSA), whichty facilitates discovery of fitness-associated alterations and genes from SCCN lineage trees. MEDALT appears more accurate than phylogenetics approaches in reconstructing copy number lineage. From data from 20 triple-negative breast cancer patients, our approaches effectively prioritize genes that are essential for breast cancer cell fitness and predict patient survival, including those implicating convergent evolution.The source code of our study is available at https://github.com/KChen-lab/MEDALT.

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