scholarly journals Improvement of Opal Multiplex Immunofluorescence Workflow for Human Tissue Sections

2021 ◽  
Vol 69 (5) ◽  
pp. 339-346
Author(s):  
Marcella Willemsen ◽  
Gabrielle Krebbers ◽  
Marcel W. Bekkenk ◽  
Marcel B.M. Teunissen ◽  
Rosalie M. Luiten
Keyword(s):  
Human Tissue ◽  
Staining Intensity ◽  
Tissue Sections ◽  
Multiple Biomarkers ◽  
Multiple Treatments ◽  
Secondary Antibodies ◽  
Low Cell Density ◽  
Differential Degree ◽  
Different Origin

The Opal multiplex technique is an established methodology for the detection of multiple biomarkers in one section. The protocol encompasses iterative single stainings and heating-mediated removal of the primary and secondary antibodies after each staining round, leaving untouched the Opal fluorophores which are deposited onto the antigen of interest. According to our experience, repetitive heating of skin sections often results in tissue damage, indicating an urgent need for milder alternatives to strip immunoglobulins. In this study, we demonstrate that considerable heating-related damage was found not only in skin but also in tissues of different origin, mostly characterized by low cell density. Importantly, the morphology remained fully intact when sections were repetitively exposed to β-mercaptoethanol-containing stripping buffer instead of multiple heating cycles. However, target epitopes appeared sensitive at a differential degree to multiple treatments with stripping buffer, as shown by loss in staining intensity, but in all cases, the staining intensity could be restored by increment of the primary antibody concentrations. Application of β-mercaptoethanol-containing stripping buffer instead of heating for antibody removal markedly improved the quality of the Opal multiplex technique, as a substantial higher number of differently colored cells could be visualized within a well-conserved morphological context:

scholarly journals Deep learning-based transformation of H&E stained tissues into special stains

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kevin de Haan ◽  
Yijie Zhang ◽  
Jonathan E. Zuckerman ◽  
Tairan Liu ◽  
Anthony E. Sisk ◽  
...  
Keyword(s):  
Visual Inspection ◽  
Kidney Diseases ◽  
Periodic Acid ◽  
Needle Core Biopsy ◽  
Silver Stain ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Biopsy Tissue ◽  
Hematoxylin And Eosin

AbstractPathology is practiced by visual inspection of histochemically stained tissue slides. While the hematoxylin and eosin (H&E) stain is most commonly used, special stains can provide additional contrast to different tissue components. Here, we demonstrate the utility of supervised learning-based computational stain transformation from H&E to special stains (Masson’s Trichrome, periodic acid-Schiff and Jones silver stain) using kidney needle core biopsy tissue sections. Based on the evaluation by three renal pathologists, followed by adjudication by a fourth pathologist, we show that the generation of virtual special stains from existing H&E images improves the diagnosis of several non-neoplastic kidney diseases, sampled from 58 unique subjects (P = 0.0095). A second study found that the quality of the computationally generated special stains was statistically equivalent to those which were histochemically stained. This stain-to-stain transformation framework can improve preliminary diagnoses when additional special stains are needed, also providing significant savings in time and cost.

Productive and reproductive traits of first-calf heifers of different origin under the conditions in LLC “Green lines – Kaluga”

2021 ◽  
pp. 3-14
Author(s):  
N. M. Kostomakhin ◽  
T. N. Pimkina ◽  
O. A. Voronkova ◽  
M. A. Gabedava ◽  
E. G. Cheremukha
Keyword(s):  
Reproductive Traits ◽  
Industrial Complex ◽  
Milk Productivity ◽  
Cattle Breeding ◽  
Linear Selection ◽  
Dairy Cattle Breeding ◽  
Line Breeding ◽  
Kaluga Region ◽  
Different Origin

Dairy cattle breeding have always occupied and will continue to occupy one of the leading places in the agro-industrial complex in our country. The most eff ective method of improving existing breeds according to opinion of a number of scientists is selection in line breeding. At the same time the evaluation of breeding sires by the quality of offspring is the cornerstone of breeding work. The purpose of the research was to analyze the influence of breeding sires, as well as their linear affiliation on the economically useful characteristics of first-calf heifers under specific conditions of maintenance and use. An analysis of the milk productivity and reproductive traits of first-calf heifers of different origin by sires and different line belonging has been presented in the article. The results of intra-linear selection and crosses of genealogical lines and their influence on the productive and reproductive traits of first-calf heifers have been evaluated. The research was carried out in LLC “Green Lines-Kaluga” of the Kaluga region. It has been established that for fixing in the genotype of the offspring of plentiful milk and fat milk in purebred breeding, it is necessary to use the following sires: Aragon 2102, Tabor 1292, Lira 2047, Phlox 1448, Chudny 1167, Yalik 397, Tabun 1277, as well as intra-linear selection in the line of Montwick Chieftain 9567929 and crosses of lines in combinations: Siling Traijun Rocket 252803 – Reflection Sovereign 198998 and Wes Ideal 933122 – Siling Traijun Rocket 252803. In order to reduce the age of the first insemination of maternal stock we recommend using a combination of lines: Wes Ideal 933122 – Montwick Chieftain 9567929, Wes Ideal 933122 – Reflection Sovereign 198998, Montwick Chieftain 9567929 – Wes Ideal 933122, Montwick Chieftain 9567929 – Reflection Sovereign 198998 and Siling Trijun Rocket 252803 – Wes Ideal 933122.

scholarly journals STUDIES ON NUCLEIC ACID METACHROMASY

1965 ◽  
Vol 27 (2) ◽  
pp. 327-336 ◽  
Author(s):  
Ned Feder ◽  
Merrill K. Wolf
Keyword(s):  
Nucleic Acid ◽  
Ribosomal Rna ◽  
Toluidine Blue ◽  
Color Contrast ◽  
Tissue Sections ◽  
Dna And Rna ◽  
Excellent Preservation ◽  
Structural Preservation ◽  
Intense Color

Acrolein-fixed, polyester wax-embedded tissue sections showed excellent preservation of light microscopic architecture and, when stained with toluidine blue, intense color contrast between DNA, which stained orthochromatically, and RNA, which stained metachromatically. This method has practical value for differentiating DNA from RNA in the same section. The color contrast was impaired by substituting formaldehyde for acrolein or paraffin for polyester wax, and was negligible in tissues fixed in formaldehyde or Carnoy's fluid and embedded in paraffin. Quality of structural preservation paralleled degree of color contrast. Metachromatic staining can be analysed, by the quantitative parameters of Bradley and colleagues, to provide inferences regarding the conformation of biopolymers in tissue sections. Comparison of the nucleic acid color contrasts in toluidine blue-stained sections with titrations of fixative-treated nucleic acids against toluidine blue in solution indicated a greater difference in conformation between DNA- and RNA-protein in acrolein-polyester sections than between acrolein-treated free DNA and RNA in solution. This is supported by recent evidence that the conformation of ribosomal RNA is quite different in whole ribosomes from that assumed by the same RNA free in solution. The acrolein-polyester method may enhance color contrast by providing superior preservation of ordered nucleoprotein conformations.

Multiple Iteractive Labeling by Antibody Neodeposition (MILAN)

2019 ◽  
Author(s):  
Giorgio Cattoretti ◽  
Francesca Maria Bosisio ◽  
Lukas Marcelis ◽  
Maddalena Maria Bolognesi
Keyword(s):  
Image Analysis ◽  
Tissue Section ◽  
Indirect Immunofluorescence ◽  
Analysis Software ◽  
Tissue Sections ◽  
Image Analysis Software ◽  
Single Section ◽  
Fluorescent Image ◽  
Secondary Antibodies

Abstract Multiplexing, labeling for multiple immunostains the very same cell or tissue section in situ, is of considerable interest. The major obstacles to the diffusion of this technique are high costs in custom antibodies and instruments, low throughput, scarcity of specialized skills or facilities. We have validated and detail here a method based on common primary and secondary antibodies, diffusely available fluorescent image scanners and routinely processed tissue sections \(FFPE). It entails rounds of four-color indirect immunofluorescence, image acquisition and removal \(stripping) of the antibodies, before another stain is applied. The images are digitally registered and the autofluorescence is subtracted. Removal of antibodies is accomplished by disulphide cleavage. In excess of 50 different antibody stains can be applied to one single section from routinely fixed and embedded tissue. This method requires a modest investment in hardware and materials and uses freeware image analysis software.

Traffic Controller for Handling Service Quality in Multimedia Network

2010 ◽  
pp. 96-112
Author(s):  
Manjunath Ramachandra ◽  
Vikas Jain
Keyword(s):  
Real Time ◽  
Long Range ◽  
Multimedia Data ◽  
Time Data ◽  
Self Similarity ◽  
Real Time Data ◽  
Data Transfers ◽  
Multimedia Network ◽  
Different Origin

The present day Internet traffic largely caters for the multimedia traffic throwing open new and unthinkable applications such as tele-surgery. The complexity of data transactions increases with a demand for in time and real time data transfers, demanding the limited resources of the network beyond their capabilities. It requires a prioritization of data transfers, controlled dumping of data over the network etc. To make the matter worse, the data from different origin combine together imparting long lasting detrimental features such as self similarity and long range dependency in to the traffic. The multimedia data fortunately is associated with redundancies that may be removed through efficient compression techniques. There exists a provision to control the compression or bitrates based on the availability of resources in the network. The traffic controller or shaper has to optimize the quality of the transferred multimedia data depending up on the state of the network. In this chapter, a novel traffic shaper is introduced considering the adverse properties of the network and counteract with the same.

scholarly journals Pain Relief as an Integral Part of the Palliative Care

2018 ◽  
Vol 6 (4) ◽  
pp. 739-741 ◽  
Author(s):  
Marija Sholjakova ◽  
Vesna Durnev ◽  
Andrijan Kartalov ◽  
Biljana Kuzmanovska
Keyword(s):  
Quality Of Life ◽  
Palliative Care ◽  
Pain Relief ◽  
Pain Control ◽  
Palliative Therapy ◽  
The Other ◽  
Spiritual Problems ◽  
Active Care ◽  
Different Origin

BACKGROUND: Palliative therapy represents active care for patients whose illness has such nature that is not responding to the curative treatment.  The palliative care aims to provide comfort and prevention from the suffering of the patients at the end of their life. Treatment of the pain presents an important integral part of palliative care.AIM: This article aims to discuss and answer to some of the analgesic regimes and therapeutic dilemmas.RESULTS: Pain control, in addition to the other treatments such as alleviation of psychological, sociological and spiritual problems, has a priority. The proper pain management can achieve a better quality of life for the patients and their families.CONCLUSION: It can be concluded that because of the different origin of the pain, the use of analgesic therapy should be individualised and adapted to the real need of every person. Finally, only a good organisation and institutionalisation of the palliative care in one society could permit better prevention of suffering at the end of the life.

scholarly journals A study examining the effects of tissue processing on human tissue sections using vibrational spectroscopy

2005 ◽  
Vol 38 (1-2) ◽  
pp. 121-127 ◽  
Author(s):  
Eoghan Ó Faoláin ◽  
Mary B. Hunter ◽  
Joe M. Byrne ◽  
Peter Kelehan ◽  
Mary McNamara ◽  
...  
Keyword(s):  
Vibrational Spectroscopy ◽  
Human Tissue ◽  
Tissue Processing ◽  
Tissue Sections
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