scholarly journals A reliable method for simultaneous demonstration of two antigens using a novel combination of immunogold-silver staining and immunoenzymatic labeling.

1990 ◽  
Vol 38 (3) ◽  
pp. 307-313 ◽  
Author(s):  
R Gillitzer ◽  
R Berger ◽  
H Moll
Keyword(s):  
Alkaline Phosphatase ◽  
Silver Staining ◽  
Immunohistochemical Staining ◽  
Reliable Method ◽  
Staining Technique ◽  
Staining Procedure ◽  
Specific Cell ◽  
Double Immunofluorescence ◽  
Novel Approach

We have developed a reliable and sensitive immunohistochemical staining technique which allows the simultaneous demonstration of two different antigens expressed in or on the same cell (referred to as mixed labeling), together with the evaluation of the general histopathological appearance of the tissue. The staining procedure combines a three-step (streptavidin-biotin) immunogold-silver staining (IGSS) with a three-step immunoenzymatic labeling. For this purpose, we investigated the compatibility of IGSS with various substrates of peroxidase or alkaline phosphatase (AP). Highly reliable and discernible mixed labeling was achieved only after initial labeling with IGSS followed by AP labeling using the substrates naphthol AS-MX phosphate/Fast Blue or naphthol AS-BI phosphate/New Fuchsin, respectively. To ensure utmost specificity, we applied FITC-conjugated mouse monoclonal antibodies and rabbit anti-FITC immunoglobulins visualized by AP-labeled immunoglobulins and the respective substrate in a final step. This novel approach provides an excellent means for demonstration of immunocompetent cells and unequivocal determination of the percentage of specific cell subsets in infiltrated tissue. The advantages of this method, as compared with double immunofluorescence or double immunoenzymatic labeling, were investigated and are discussed.

scholarly journals Simultaneous demonstration of lectin-binding sites and antigens detected by monoclonal antibodies in a parallelized double-staining technique: a highly discriminative and quickly developing technique for frozen sections.

1986 ◽  
Vol 34 (3) ◽  
pp. 399-402 ◽  
Author(s):  
G Mechtersheimer ◽  
P Möller ◽  
F Momburg ◽  
G Moldenhauer ◽  
K Schwechheimer
Keyword(s):  
Alkaline Phosphatase ◽  
Monoclonal Antibodies ◽  
Binding Sites ◽  
Lectin Binding ◽  
Staining Technique ◽  
Staining Procedure ◽  
Double Staining ◽  
Enzyme Complexes ◽  
Simultaneous Visualization ◽  
Lectin Binding Sites

A sensitive immunoenzymatic double-staining technique is presented for the simultaneous visualization of lectin-binding sites and antigenic structures detected by monoclonal antibodies. The lectin is demonstrated by an extended unlabeled peroxidase-antiperoxidase (PAP) technique and the monoclonal antibody by an alkaline phosphatase-antialkaline phosphatase (APAAP) method, which corresponds to the standard PAP technique. 3-amino-9-ethylcarbazole (AEC) and fast blue BB salt serve as substrates for the peroxidase and the alkaline phosphatase, respectively. The antisera and the enzyme complexes raised in different animals enable the performance of three parallel incubation steps. The staining procedure requires three and a half hours altogether. This method proved to be highly discriminative and rather insensitive to interference by various artifacts.

The Use of TMAH to Etch Silicon and Expose Metal Bridging Failures

2000 ◽  
Author(s):  
Mark Morris ◽  
James Mohr ◽  
Esteban Ortiz ◽  
Steven Englebretson
Keyword(s):  
Failure Mechanism ◽  
Schottky Diodes ◽  
Tetramethylammonium Hydroxide ◽  
Novel Approach ◽  
Failure Location ◽  
Metal Etching ◽  
Plastic Mold

Abstract Determination of metal bridging failures on plastic encapsulated devices is difficult due to the metal etching effects that occur while removing many of the plastic mold compounds. Typically, the acids used to remove the encapsulation are corrosive to the metals that are found within the device. Thus, decapsulation can result in removal of the failure mechanism. Mechanical techniques are often not successful due to damage that results in destruction of the die and failure mechanism. This paper discusses a novel approach to these types of failures using a silicon etch and a backside evaluation. The desirable characteristics of the technique would be to remove the silicon and leave typical device metals unaffected. It would also be preferable that the device passivation and oxides not be etched so that the failure location is not disturbed. The use of Tetramethylammonium Hydroxide (TMAH), was found to fit these prerequisites. The technique was tested on clip attached Schottky diodes that exhibited resistive shorting. The use of the TMAH technique was successful at exposing thin solder bridges that extruded over the edge of the die resulting in failure.

The Need for Using An Enzymatic Colorimetric Assay in Creatinine Determination of Peritoneal Dialysis Solutions

1990 ◽  
Vol 10 (1) ◽  
pp. 89-92 ◽  
Author(s):  
Liliane Larpent ◽  
Christian Verger
Keyword(s):  
Peritoneal Dialysis ◽  
Reliable Method ◽  
Colorimetric Assay ◽  
Colorimetric Method ◽  
Peritoneal Membrane ◽  
Creatinine Kinetics ◽  
Dialysis Solutions ◽  
Peritoneal Dialysis Solutions ◽  
Enzymatic Test

The fate of the peritoneal membrane on continuous ambulatory peritoneal dialysis (CAPD) is usually evaluated through the modification of its permeability to various solutes as glucose, creatinine, and urea. Therefore, the accuracy of the methods used for measurements of creatinine is of great importance. A particular problem does exist for creatinine determination as it may be influenced by the presence of glucose. We studied a new enzymatic colorimetric method for creatinine determination in peritoneal dialysis solutions which contain high dextrose concentrations. Creatinine was measured in plasma, urine, and dialysate from 18 patients on CAPD and in pure dextrose solutions, with an enzymatic test (Boehringer Mannheim) and with Jaffe's reaction on two different analyzers: Astra (Beckman) and Eris (Merck). Creatinine results were similar with both assays (Jaffe's reaction and enzymatic test) when measured in blood and urine. However the Jaffe's reaction gave higher creatinine results than the enzymatic test (p < 0.001), when assays were performed in peritoneal dialysis solutions and in pure glucose solutions. In addition, it appeared that other components of dialysis solutions, mainly calcium chloride, influenced unpredictably the results of creatinine with the Jaffe's reaction. We conclude that specific enzymatic test is a more accurate and reliable method to evaluate creatinine kinetics through the peritoneal membrane when determined in CAPD solutions.

par-4, a gene required for cytoplasmic localization and determination of specific cell types in Caenorhabditis elegans embryogenesis.

Genetics ◽  
1992 ◽  
Vol 130 (4) ◽  
pp. 771-790 ◽  
Author(s):  
D G Morton ◽  
J M Roos ◽  
K J Kemphues
Keyword(s):  
Caenorhabditis Elegans ◽  
Cell Types ◽  
Intestinal Cells ◽  
Specific Cell ◽  
Cytoplasmic Localization ◽  
Loss Of Function ◽  
Embryo Viability ◽  
Cell Stage ◽  
Maternal Genome

Abstract Specification of some cell fates in the early Caenorhabditis elegans embryo is mediated by cytoplasmic localization under control of the maternal genome. Using nine newly isolated mutations, and two existing mutations, we have analyzed the role of the maternally expressed gene par-4 in cytoplasmic localization. We recovered seven new par-4 alleles in screens for maternal effect lethal mutations that result in failure to differentiate intestinal cells. Two additional par-4 mutations were identified in noncomplementation screens using strains with a high frequency of transposon mobility. All 11 mutations cause defects early in development of embryos produced by homozygous mutant mothers. Analysis with a deficiency in the region indicates that it33 is a strong loss-of-function mutation. par-4(it33) terminal stage embryos contain many cells, but show no morphogenesis, and are lacking intestinal cells. Temperature shifts with the it57ts allele suggest that the critical period for both intestinal differentiation and embryo viability begins during oogenesis, about 1.5 hr before fertilization, and ends before the four-cell stage. We propose that the primary function of the par-4 gene is to act as part of a maternally encoded system for cytoplasmic localization in the first cell cycle, with par-4 playing a particularly important role in the determination of intestine. Analysis of a par-4; par-2 double mutant suggests that par-4 and par-2 gene products interact in this system.

Urinary gonadotropin measurements by enhanced luminometric assays (LIA) for the evaluation of pubertal development

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
And Demir ◽  
Adem Aydın ◽  
Atilla Büyükgebiz ◽  
Ulf-Håkan Stenman ◽  
Matti Hero
Keyword(s):  
Healthy Subjects ◽  
Reliable Method ◽  
Pubertal Development ◽  
High Sensitivity ◽  
Tanner Stage ◽  
Time Resolved ◽  
Stage 1 ◽  
Sensitive Detection Technique ◽  
Serum And Urine

Abstract Objectives Determination of LH in urine has proved to be a reliable method for evaluation of pubertal development. The human LH assay based on time-resolved immunofluorometric (IFMA) technology (AutoDELFIA, PerkinElmer, Wallac) has been found to be suitable for this purpose thanks to its high sensitivity but other assays have not been evaluated. We have analyzed our data obtained by another potentially sensitive detection technique, enhanced luminometric assay (LIA) with the objective of finding a viable alternative to IFMA since these may not be available in the future. Methods LIA was used to measure LH and FSH in serum and urine samples from 100 healthy subjects of each Tanner stage and both genders, whose pubertal development has been determined. Results Urinary gonodotropin concentrations measured by LIA correlated well with Tanner stage [(r=0.93 for girls, r=0.81 for boys; p<0.01 for LH) and (r=0.81 for girls, r=0.73 for boys; p<0.01 for FSH)]. LIA determinations revealed the increase in U-LH concentrations during the transition from Tanner stage 1–2 in both girls and boys (p<0.001), whereas U-FSH and S-LH were able to detect the increase from Tanner stage 1–2 only in boys or girls, respectively (both p<0.001). Conclusions Measurement of urinary gonadotropin concentrations by LIA may be useful for the evaluation of overall pubertal development and also in the detection of transition from prepuberty to puberty.

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