Transmission electron microscopy (TEM)- by sivasangari Shanmugam. Transmission electron microscopy (TEM) is a technique used to observe the features of very small specimens.
A scanning electron microscope is a type of electron microscope that produces images of a sample by scanning the surface with a focused beam of electrons. The electrons interact with atoms in the sample, producing various signals that contain information about the sample's surface topography and composition.
SEMs can magnify an object from about 10 times up to 300,000 times. A scale bar is often provided on an SEM image. From this the actual size of structures in the image can be calculated.
Transmission electron microscopy (TEM)- by sivasangari Shanmugam. Transmission electron microscopy (TEM) is a technique used to observe the features of very small specimens.
A scanning electron microscope is a type of electron microscope that produces images of a sample by scanning the surface with a focused beam of electrons. The electrons interact with atoms in the sample, producing various signals that contain information about the sample's surface topography and composition.
SEMs can magnify an object from about 10 times up to 300,000 times. A scale bar is often provided on an SEM image. From this the actual size of structures in the image can be calculated.
SEM is a type of electron microscope designed for directly studying the surfaces of solid objects, that utilizes a beam of focused electron of relatively low energy as an electron probe that is scanned in a regular manner over the specimen.
Electron microscope, principle and applicationKAUSHAL SAHU
Introduction
History
Resolution &Magnification of
Electron microscope
Types of electron microscope
1) Transmission electron microscope (TEM)
- Structural parts of TEM
- Principle & Working of TEM
- Sample preparation for TEM
- Advantages & disadvantages of TEM
Scanning electron microscope (SEM)
- Structural parts of SEM
- Principle & Working of SEM
- Sample preparation for SEM
- Advantages & disadvantages of SEM
3) Scanning transmission electron microscope (STEM)
Applications of electron microscope
Conclusion
References
SEM is a technique that provides information such as topography, composition and crystallographic information of an object.
Scanning electron microscopes use a beam of highly energetic electrons to examine objects on a very fine scale.
SEM produces images by detecting secondary electrons that are emitted from the surface due to excitation from a primary electron beam.
The TEM is a very powerful tool for material science.
TEM can be used to study the growth of layers, their composition and defects in semiconductors.
High resolution can be used to analyze the quality, shape, size and density of quantum wells, wires and dots.
This presentation include information about electron microscope & types of electron microscope i.e. SEM (Scanning electron microscope) & TEM (Transmission electron microscope).
An electron microscope is a microscope that uses a beam of scattered electrons as a source of illumination. It is used to get information about structure, topology, morphology & composition of materials. It has many advantages. Basically there are 4 types of electron microscope but here we will discuss only 2 types.
Transmission electron microscopy is a microscopy technique in which a beam of electrons is transmitted through an ultra-thin specimen, interacting with the specimen as it passes through it. Its resolution & magnification is about 10,000,000x. There are 5 types of transmission electron microscope i.e. BFTEM (Bright field transmision electron microscope), DFTEM (Dark field transmission electron microscope), HRTEM (High resolution transmission electron microscope), EFTEM (Energy filtered transmission electron microscope), ED (Electron diffraction). there are 4 techniques of TEM i.e. negative staining, shadow casting, Freeze fracture replication, freeze etching. It has many applications e.g, for the study of Cancer research, virology, chemical industry, electronic structure etc.
A scanning electron microscope is a type of electron microscope that produces images of a sample by scanning it with a focused beam of electrons. Types of signals produce by SEM include secondary electrons, back scattered electrons, X-rays, light rays. There are many advantages of SEM e.g, Btter resolution, fast imaging easy to operate, work with low voltage etc.
5. Microsocope ELECTRON MICROSCOPE (TEM & SEM ) - BasicsNethravathi Siri
Basics only
Electron beam is the source of illumination.
Image is produced by magnetic field.
Contrasting features between light microscope and electron microscope are
construction, working principle, specimen preparation, cost-expenses and designed
room (vacuum chamber).
A scanning electron microscope (SEM) is a type of electron microscope that produces images of a sample by scanning the surface with a focused beam of electrons.
SEM is a type of electron microscope designed for directly studying the surfaces of solid objects, that utilizes a beam of focused electron of relatively low energy as an electron probe that is scanned in a regular manner over the specimen.
Electron microscope, principle and applicationKAUSHAL SAHU
Introduction
History
Resolution &Magnification of
Electron microscope
Types of electron microscope
1) Transmission electron microscope (TEM)
- Structural parts of TEM
- Principle & Working of TEM
- Sample preparation for TEM
- Advantages & disadvantages of TEM
Scanning electron microscope (SEM)
- Structural parts of SEM
- Principle & Working of SEM
- Sample preparation for SEM
- Advantages & disadvantages of SEM
3) Scanning transmission electron microscope (STEM)
Applications of electron microscope
Conclusion
References
SEM is a technique that provides information such as topography, composition and crystallographic information of an object.
Scanning electron microscopes use a beam of highly energetic electrons to examine objects on a very fine scale.
SEM produces images by detecting secondary electrons that are emitted from the surface due to excitation from a primary electron beam.
The TEM is a very powerful tool for material science.
TEM can be used to study the growth of layers, their composition and defects in semiconductors.
High resolution can be used to analyze the quality, shape, size and density of quantum wells, wires and dots.
This presentation include information about electron microscope & types of electron microscope i.e. SEM (Scanning electron microscope) & TEM (Transmission electron microscope).
An electron microscope is a microscope that uses a beam of scattered electrons as a source of illumination. It is used to get information about structure, topology, morphology & composition of materials. It has many advantages. Basically there are 4 types of electron microscope but here we will discuss only 2 types.
Transmission electron microscopy is a microscopy technique in which a beam of electrons is transmitted through an ultra-thin specimen, interacting with the specimen as it passes through it. Its resolution & magnification is about 10,000,000x. There are 5 types of transmission electron microscope i.e. BFTEM (Bright field transmision electron microscope), DFTEM (Dark field transmission electron microscope), HRTEM (High resolution transmission electron microscope), EFTEM (Energy filtered transmission electron microscope), ED (Electron diffraction). there are 4 techniques of TEM i.e. negative staining, shadow casting, Freeze fracture replication, freeze etching. It has many applications e.g, for the study of Cancer research, virology, chemical industry, electronic structure etc.
A scanning electron microscope is a type of electron microscope that produces images of a sample by scanning it with a focused beam of electrons. Types of signals produce by SEM include secondary electrons, back scattered electrons, X-rays, light rays. There are many advantages of SEM e.g, Btter resolution, fast imaging easy to operate, work with low voltage etc.
5. Microsocope ELECTRON MICROSCOPE (TEM & SEM ) - BasicsNethravathi Siri
Basics only
Electron beam is the source of illumination.
Image is produced by magnetic field.
Contrasting features between light microscope and electron microscope are
construction, working principle, specimen preparation, cost-expenses and designed
room (vacuum chamber).
A scanning electron microscope (SEM) is a type of electron microscope that produces images of a sample by scanning the surface with a focused beam of electrons.
It is technique used to identify more clear picture of small organelles which are not able to distinguished by normal microscopy techniques. Its too expensive to perform that's why not used commonly.
Transmission Electron Microscope (TEM), RESOLVING POWER, Scanning Electron Microscope, PRINCIPLE AND WORKING OF SEM, SEM SAMPLE PREPARATION, Limitations of Scanning Electron Microscopy (SEM), ADVANTAGES & DISADVANTAGES OF SEM, APPLICATIONS OF SEM, PRINCIPLE, AND WORKING OF TEM, SAMPLE PREPARATION FOR TEM, ADVANTAGES & DISADVANTAGES OF TEM, APPLICATIONS OF TEM, Differences between SEM and TEM.
Electron Microscopy - Scanning electron microscope, Transmission Electron Mic...Sumer Pankaj
An electron microscope is a microscope that uses a beam of accelerated electrons as a source of illumination. As the wavelength of an electron can be up to 100,000 times shorter than that of visible light photons, electron microscopes have a higher resolving power than light microscopes and can reveal the structure of smaller objects. A transmission electron microscope can achieve better than 50 pm resolution and magnifications of up to about 10,000,000x whereas most light microscopes are limited by diffraction to about 200 nm resolution and useful magnifications below 2000x.
Electron microscopes are used to investigate the ultrastructure of a wide range of biological and inorganic specimens including microorganisms, cells, large molecules, biopsy samples, metals, and crystals. Industrially, electron microscopes are often used for quality control and failure analysis. Modern electron microscopes produce electron micrographs using specialized digital cameras and frame grabbers to capture the image.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
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.
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.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
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.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
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.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
2. 1- Introduction to Microscopy
2- Electron Microscope
History of Electron Microscope
Types of electron Microscope
3- Transmission Electron Microscope
History of TEM
Components of TEM
Sample Preparation in TEM
Working of TEM
Advantages of TEM
Limitation of TEM
Applications of TEM
4- Scanning Electron Microscope
History of SEM
Components of SEM
Sample Preparation in SEM
Working of SEM
Advantages of SEM
Limitation of SEM
Applications of SEM
4- Difference between Light and Electron
Microscope.
5- Difference between TEM & SEM.
3. An optical instrument used for viewing very small objects, such as
mineral samples or animal or plant cells, typically magnified several
hundred times.
The branch of science that deals with the study of the construction,
working principles and use of microscopes.
4. An electron microscope is a type of microscope that uses an
electron beam to illuminate a specimen and produce a
magnified image.
Advantages of electron microscope over light microscope :
An EM has greater resolving power than ordinary one.
Can reveal the structure of smaller objects because electrons have
wavelengths about 100,000 times shorter than visible light .
They can achieve magnification of up to about 10,000,000x whereas
ordinary, light microscopes are about 200 nm resolution and useful
magnifications below 2000x.
5. HISTORY.
Hertz (1857-94) suggested that cathode rays were a form of wave motion .
It had earlier been recognized by Plücker in 1858 that the deflection of "cathode
rays" (electrons) was possible by the use of magnetic fields. This effect had been
utilized to build primitive cathode ray oscilloscopes (CROs) as early as 1897 by
Ferdinand Braun, intended as a measurement device.
Weichert, in 1899, found that these rays could be concentrated into a small spot by
the use of an axial magnetic field produced by a long solenoid.
In 1926 that Busch showed theoretically that a short solenoid converges a beam of
electrons in the same way that glass can converge the light of the sun.
Busch should probably therefore be known as the father of electron optics.
6. 1. Transmission electron microscope (TEM)
2. Scanning electron microscope (SEM)
3. Reflection electron microscope (REM)
4. Scanning transmission electron microscope (STEM)
7.
8. Ernst Ruska developed the first
electron microscope, a TEM,
with the assistance of Max
Knolls in 1931.
After significant improvements
to the quality of magnification,
Ruska joined the Sieman’s
Company in the late 1930s as
an electrical engineer, where he
assisted in the manufacturing
of his TEM
The first commercial TEM
was available in 1939.
10. From the top down, the TEM
consists of an emission source,
which may be a
, or a
source.
For tungsten, this will be of
the form of either a hairpin-
style filament, or a small spike-
shaped filament and for LaB6
sources utilize small single
crystals.
By connecting this gun to a
high voltage source (typically
~100–300 kV) the gun will,
begin to emit electrons.
11. are the first in the
series. They are also called the
"roughing pumps" as they are used to
initially lower the pressure within the
column through which the electron
must travel to 10 -3 mm of Hg range.
may achieve
higher vacuums (in the 10-5 mm Hg
range) but must be backed by the
rotary pump.
In addition a
, when an even greater
vacuum is required.
12. Electromagnetic lenses are made of a
coil of copper wires inside several iron
pole pieces .
Magnetic condenser lens: -
converged the electron beam on the
specimen.
Magnetic objective lens: - Focuses
the electron into the first real image of
the object which is enlarged 2000
times.
Magnetic intermediate lens : -forms
an intermediate image of the specimen
Magnetic projector lens: - it then
magnifies a portion of the first image
13. An image is formed from the
interaction of the electrons
transmitted through the
specimen; the image is
magnified and focused onto an
imaging device, such as:-
A fluorescent screen
On a layer of photographic
film, or to be detected by a
sensor such as a CCD
"Charged Coupled Device"
camera.
14. The specimen holders are adapted to hold a
standard size of grid upon which the sample is
placed or a standard size of self-supporting
specimen.
Usual grid materials are copper,
molybdenum, gold or platinum. This grid is
placed into the sample holder, which is paired
with the specimen stage.
The most common is the side entry holder,
where the specimen is placed near the tip of a
long metal (brass or stainless steel) rod, with
the specimen placed flat in a small bore.
Along the rod are several polymer vacuum
rings to allow for the formation of a vacuum
seal of sufficient quality, when inserted into the
stage.
15. 1. Fixation: The first step in sample preparation, has the aim
of preserving tissue in its original state. Fixatives must
be buffered to match the pH and osmolarity of the living
tissue. Eg . Glutaraldehyde
2. Dehydration: - Before sample can be transferred to resin all
the water must be removed from the sample. This is carried
out using a graded ethanol series.
3. Tissue sectioning: -By passing samples over a glass or
diamond edge, small, thin sections can be readily obtained
using a semi-automated method.
Contd…
16. • Sample staining: - Details in light microscope
samples can be enhanced by stains that absorb light;
similarly TEM samples of biological tissues can utilize
high atomic number stains to enhance contrast. The
stain absorbs electrons or scatters part of the
electron beam Compounds of heavy metals such as
osmium, lead, uranium or gold (in immunogold
labelling) may be used
• Mechanical milling: -Mechanical polishing may be
used to prepare samples. Polishing needs to be done
to a high quality, to ensure constant sample
thickness across the region of interest. A diamond, or
cubic boron nitride polishing compound may be used
17. The electron source is commonly a tungsten filament
of 30-150 KV potential. The electron beam passes
through the centre of ring-like magnetic condenser
and becomes converged on the specimen. After being
transmitted through the specimen, the magnetic
objective focuses the electron into the first (real)
image of the object which is enlarged (2000 times).
The magnetic projector lens then magnifies a portion
of the first image producing magnification up to 240,
000 or more.
The final enlarged image can be viewed by
striking on fluorescent screen which makes it visible.
The image can also be thrown upon a photographic
plate for permanent record.
Molecules in the microscope interfere with
the movement of electrons. To prevent this, the
interior of the microscope is kept in the state of high
vacuum, around 10.4-10.6 mmHg. It is also necessary
to have specimen ultra thin. The electron beams have
a poor penetrating power, therefore, only small
objects or very thin sections of the specimen can be
examined.
18.
19. • TEMs offer the most powerful magnification, potentially over
one million times or more.
• TEMs have a wide-range of applications and can be utilized in a
variety of different scientific, educational and industrial fields.
• TEMs provide information on element and compound structure.
• Images are high-quality and detailed.
• TEMs are able to yield information of surface features, shape,
size and structure.
• They are easy to operate with proper training.
20. • TEMs are large and very expensive.
• Laborious sample preparation.
• Potential artifacts from sample preparation.
• Operation and analysis requires special training.
• Samples are limited to those that are electron
transparent, able to tolerate the vacuum chamber and
small enough to fit in the chamber.
• TEMs require special housing and maintenance.
• Images are black and white.
21. 1. Cancer research
2. Virology
3. Materials science as well as pollution
4. Nanotechnology and
5. Semiconductor research.
22.
23. The earliest known work
describing the concept of a
Scanning Electron Microscope
was by M. Knoll (1935).
In 1965 the first commercial
instrumented SEM by
Cambridge Scientific Instrument
Company as the "Stereoscan”.
The first SEM used to examine
the surface of a solid specimen
was described by Zworykin et al.
(1942).
25. Tungsten is normally used in
thermionic electron guns because it has
the highest melting point and lowest
vapour pressure of all metals, thereby
allowing it to be heated for electron
emission, and because of its low cost.
Other types of electron emitters
include lanthanum hexaboride (LaB
6) cathodes.
In a typical SEM, an electron beam is
thermionically emitted from an electron
gun fitted with a tungsten filament
cathode.
26. There are two main lenses used
in SEM: -
1. Condenser lenses
2. Objective lenses
Main role of the condenser lens
is to control the size of the beam
The objective lens focuses
electrons on the sample at the
working distance
27. A high vacuum minimises
scattering of the electron beam
before reaching the specimen.
This is important as scattering or
attenuation of the electron beam will
increase the probe size and reduce
resolution, especially in the SE
mode.
The high vacuum condition also
optimises collection efficiancy,
especially of the secondary
electrons.
28. An electron detector is
placed in the sample
chamber.
By having a 10 keV
positive potential on its face,
it attracts the secondary
electrons emitted from the
sample surface.
One advantage of this
biased detector is that it can
attract secondary electrons
emitted from sides of the
sample which are physically
blocked from the detector
face.
This greatly reduces
shadowing effects in SEM
images.
29. • Some samples, such as hard tissues like bone or
teeth, and organisms with a tough exoskeleton, such
as some arthropods, can be studied without any
preparation, but these are the exception.
• Biological specimens, such as cells and tissues or
tissue components, must first be fixed to preserve
their native structure.
• Fixation is done either by chemical or physical
means.
contd. . .
30. • Chemical fixation uses formalin or glutaraldehyde of
varying per cent concentrations in a buffer of a specific
pH and osmolarity. Physical fixation may be by heat (such
as boiling an egg), but is more commonly done by
freezing.
• Hydrated samples, like most biological and some
materials specimens, must first be dehydrated before
placing the specimen in the SEM sample chamber. This is
typically done by passing the specimens through a
graded series of ethanol-water mixtures to 100%.
• If the specimen is an electron conductor, it needs only to
be held on an appropriate support. If it is non-conductor
it is allowed to dry but if moist, freeze dried in liquid
nitrogen is necessary. The specimen is then coated with
metal vapour (gold) in vacuum.
31. The scanning electron microscope gives 3D
(dimensional surface) views of objects.
The electron originates at high energy
(20,000 v) from a hot tungsten cathode
“gun”. These electrons are sharply focused,
adjusted and narrow by an arrangement of
magnetic fields. The primary beam (Probe)
acts only as an exciter of image forming
secondary electrons emerging from the
surface of the specimen. The probe scans
the specimen. Images are elicited from
wherever the probe strikes the metal coated
areas of the specimen.
The useful secondary electrons are
magnetically deflected to a collector or
detector. The successive signal from the
collector are amplified and transmitted to a
cathode ray (T.V.) tube. The scanning beam
and T.V. tube beam are synchronized. The
image scans by the eye on T.V. screen. The
T.V. image may be photographed,
videotaped or processed in motion on a
computer.
32. 1. Its wide-array of applications.
2. The detailed three-dimensional and topographical imaging
and the versatile information gathered from different
detectors.
3. This instrument works fast.
4. Although all samples must be prepared before placed in
the vacuum chamber, most SEM samples require minimal
preparation actions.
33. • The disadvantages of a Scanning Electron Microscope start with the size and
cost.
• SEMs are expensive, large and must be housed in an area free of any possible
electric, magnetic or vibration interference.
• Maintenance involves keeping a steady voltage, currents to electromagnetic
coils and circulation of cool water.
• Special training is required to operate an SEM as well as prepare samples.
• The preparation of samples can result in artifacts. The negative impact can be
minimized with knowledgeable experience researchers being able to identify
artifacts from actual data as well as preparation skill.
• There is no absolute way to eliminate or identify all potential artifacts.
40. Electron Microscopy and Analysis by : P.J. GOODHEW, University
of surrey and F.J.Humphreys, UK imperial College, London, UK
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