Immunostaining is an antibody-based method to detect specific proteins in samples. It involves using antibodies that are conjugated to detectable labels like fluorescence or enzymatic tags. There are different immunostaining methods based on the label used, such as immunofluorescence, immunocolloidal gold, or immunoperoxidase staining. Procedures can involve direct or indirect staining with primary and secondary antibodies. Immunostaining is used clinically in pathology and in research to investigate protein expression, localization, and changes.
wo-dimensional gel electrophoresis, abbreviated as 2-DE or 2-D electrophoresis, is a form of gel electrophoresis commonly used to analyze proteins. Mixtures of proteins are separated by two properties in two dimensions on 2D gels. 2-DE was first independently introduced by O'Farrell and Klose in 1975.
wo-dimensional gel electrophoresis, abbreviated as 2-DE or 2-D electrophoresis, is a form of gel electrophoresis commonly used to analyze proteins. Mixtures of proteins are separated by two properties in two dimensions on 2D gels. 2-DE was first independently introduced by O'Farrell and Klose in 1975.
Gel electrophoresis native, denaturing&reducingLovnish Thakur
Electrophoresis is a technique used to separate and sometimes purify macromolecules - especially proteins and nucleic acids - that differ in size, charge or conformation.
Radial immunodiffusion (RID) or Mancini method is also known as Single radial immunodiffusion. An immunodiffusion technique, used in immunology to determine the quantity or concentration of an antigen in a sample.
Fluorescence- Activated Cell Sorter is a powerful technique used in cell sorting, cell-cycle analysis etc.
The presentation gives a basic understanding of the principle of FACS, instrumentation, interpretation of results, applications, how to do cell-cycle analysis using FACS and various troubleshooting tips.
Ouchterlony double immunodiffusion (also known as passive double immunodiffusion) is an immunological technique used in the detection, identification and quantification of antibodies and antigens, such as immunoglobulins and extractable nuclear antigens.
Immunohistochemistry (IHC) is the localization of a known antigen in tissues by utilizing antibodies directed towards that (specific) antigen. In this presentation, we will introduce the procedure of IHC and the troubleshooting solutions.
Gel electrophoresis native, denaturing&reducingLovnish Thakur
Electrophoresis is a technique used to separate and sometimes purify macromolecules - especially proteins and nucleic acids - that differ in size, charge or conformation.
Radial immunodiffusion (RID) or Mancini method is also known as Single radial immunodiffusion. An immunodiffusion technique, used in immunology to determine the quantity or concentration of an antigen in a sample.
Fluorescence- Activated Cell Sorter is a powerful technique used in cell sorting, cell-cycle analysis etc.
The presentation gives a basic understanding of the principle of FACS, instrumentation, interpretation of results, applications, how to do cell-cycle analysis using FACS and various troubleshooting tips.
Ouchterlony double immunodiffusion (also known as passive double immunodiffusion) is an immunological technique used in the detection, identification and quantification of antibodies and antigens, such as immunoglobulins and extractable nuclear antigens.
Immunohistochemistry (IHC) is the localization of a known antigen in tissues by utilizing antibodies directed towards that (specific) antigen. In this presentation, we will introduce the procedure of IHC and the troubleshooting solutions.
Immunohistochemistry (IHC) combines histological, immunological, and biochemical techniques for the identification of specific tissue components by means of a particular antigen/antibody reaction tagged with a visible label
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
The increased availability of biomedical data, particularly in the public domain, offers the opportunity to better understand human health and to develop effective therapeutics for a wide range of unmet medical needs. However, data scientists remain stymied by the fact that data remain hard to find and to productively reuse because data and their metadata i) are wholly inaccessible, ii) are in non-standard or incompatible representations, iii) do not conform to community standards, and iv) have unclear or highly restricted terms and conditions that preclude legitimate reuse. These limitations require a rethink on data can be made machine and AI-ready - the key motivation behind the FAIR Guiding Principles. Concurrently, while recent efforts have explored the use of deep learning to fuse disparate data into predictive models for a wide range of biomedical applications, these models often fail even when the correct answer is already known, and fail to explain individual predictions in terms that data scientists can appreciate. These limitations suggest that new methods to produce practical artificial intelligence are still needed.
In this talk, I will discuss our work in (1) building an integrative knowledge infrastructure to prepare FAIR and "AI-ready" data and services along with (2) neurosymbolic AI methods to improve the quality of predictions and to generate plausible explanations. Attention is given to standards, platforms, and methods to wrangle knowledge into simple, but effective semantic and latent representations, and to make these available into standards-compliant and discoverable interfaces that can be used in model building, validation, and explanation. Our work, and those of others in the field, creates a baseline for building trustworthy and easy to deploy AI models in biomedicine.
Bio
Dr. Michel Dumontier is the Distinguished Professor of Data Science at Maastricht University, founder and executive director of the Institute of Data Science, and co-founder of the FAIR (Findable, Accessible, Interoperable and Reusable) data principles. His research explores socio-technological approaches for responsible discovery science, which includes collaborative multi-modal knowledge graphs, privacy-preserving distributed data mining, and AI methods for drug discovery and personalized medicine. His work is supported through the Dutch National Research Agenda, the Netherlands Organisation for Scientific Research, Horizon Europe, the European Open Science Cloud, the US National Institutes of Health, and a Marie-Curie Innovative Training Network. He is the editor-in-chief for the journal Data Science and is internationally recognized for his contributions in bioinformatics, biomedical informatics, and semantic technologies including ontologies and linked data.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...
Immunostaining
1. IMMUNOSTAINING
SOUVIK BISWAS
M.PHARM (PHARMACOLOGY), 1ST YEAR (2ND SEM)
ROLL NO. 27720216003
REGISTRATION NO.162772310018
UNDER THE GUIDANCE OF DR. SANDIPAN DASGUPTA
NSHM KNOWLEDGE CAMPUS, KOLKATA – GROUP OF INSTITUTION
2. ANTIGENS
Antigen is any substance (such as
an immunogen or a hapten) foreign
to the body that evokes an immune
response either alone or after
forming a complex with a larger
molecule (such as a protein) and
that is capable of binding with a
product (such as an antibody or T
cell) of the immune response.
3. ANTIBODIES (IMMUNOGLOBULINS; IG)
It is a large number of proteins of high
molecular weight that are produced
normally by specialized B cells after
stimulation by an antigen and act
specifically against the antigen in an
immune response and that typically
consist of four subunits including two
heavy chains and two light chains -
called also immunoglobulin.
4. WHY DO IMMUNOSTAINING?
• Confirm genotype-phenotype
(e.g. KO, KI, levels or efficiency of expression, etc.)
• Where is a biomarker expressed?
• When is a biomarker expressed?
• Co-expression or co-localization.
• Cell type-specific biomarkers
(e.g. CD4, CD8 T-cells, NK, Macrophages, Mast Cells, etc.)
5. INTRODUCTION AND DEFINITION
• Immunostaining is a general term in
biochemistry that applies to any use of an
antibody-based method to detect a specific
protein in a sample.
• The term "immunostaining" was originally
used to refer to the immunohistochemical
staining of tissue sections, as first
described by Albert Coons in 1941.
• The key to immunohistochemistry is the
specific antibody-mediated detection of a
target antigen, known as immunostaining or
immunodetection.
6. IMMUNOSTAINING METHOD
According to different biotins conjugated with antibodies, IHC staining
methods can be classified as
• Immunofluorescence,
• Immunoenzymological staining
• Immunocolloidal gold technique
According to different kinds of procedures, immunohistochemistry staining
can be divided into subtypes of
• Direct staining (one-step staining)
• Indirect staining (two-step, three-step or multi-step staining)
• PAP staining method
7. DIFFERENT BIOTINS CONJUGATED WITH
ANTIBODIES
• Immunofluorescence:
It is the first immunohistochemical staining
method. With antigen-antibody binding reaction,
antigens are visualized by fluorescence dyes
conjugated with antibodies and that is localized
under fluorescence microscope.
• Immunoenzymological Staining:
In here enzyme-labeled antibodies are used
to bind with specific antigens in tissues samples
or cultured cells and localized by light microscope.
8. CONTD…
• Immunocolloidal Gold Technique:
It is a kind of technique that uses
colloidal gold as a marker.
Immunocolloidal gold technique is
suitable for single or multi-label
detection under immune-electron
microscope, and light microscope.
9. DIFFERENT KINDS OF PROCEDURES
• Direct Staining:
Incubate the sections with mixture of two
primary antibodies which are respectively
conjugated with two fluorescence dyes (e.g. FITC
and TRITC) Or, successively incubate sections
with two primary antibodies.
• Indirect Staining:
In here the primary antibodies are without
fluorescence dyes. Incubate sections with one
kind of primary antibody and corresponding
secondary antibody, then the other.
10. CONTD…
• PAP Staining Method (peroxidase
anti-peroxidase method):
Incubate sections with
corresponding secondary antibodies
which are conjugated with two
different enzymes (e.g. HRP, AKP), or
anti-HRP (PAP complex), anti-AKP
(APAAP complex).
12. IMMUNOHISTOCHEMISTRY STAINING TIPS
• Fresh fixed tissue
• Sufficient dehydration of tissues
• An intact, uniform and smooth sectioning
• Binder materials
• Sufficient deparaffinization
• A thorough inhibition of endogenous peroxidase
13. APPLICATIONS
• Clinically, IHC is used in histopathology for the diagnosis of
specific types of cancers based on molecular markers.
• In laboratory science, immunostaining can be used for a
variety of applications based on investigating the presence or
absence of a protein, its tissue distribution, its sub-cellular
localization, and of changes in protein expression or
degradation.
14. REFERENCES
• Formation of the first cleavage spindle in nematode embryos by Albertson,
D.GDev. Biol. 101, (1984). page no. 61–72.
• Immunohistochemistry: Methods Express, edited by Simon Renshaw, Scion
Publishing Ltd, Bloxham, UK, 1st edition (15 Dec. 2006), page no. 45-96
• http://www.abcam.com/index.html?pageconfig=resource&rid=12615
(accessed on 16.05.2017)
• http://www.mdbioproducts.com/resources/protocols/immunohistochemistr
y (accessed on 17.05.2017)
• http://www.immunohistochemistry.us/immunohistochemistry-staining.html
(accessed on 18.05.2017)