The document discusses different types of microscopes used to view microscopic specimens. It describes light microscopes, which use lenses and visible light, including brightfield, darkfield, phase contrast, and fluorescence microscopes. It also describes electron microscopes, which use electromagnetic lenses and electrons beams to view specimens, including transmission electron microscopes that pass electrons through thin specimens, and scanning electron microscopes that scan surfaces to produce 3D images. Key aspects and uses of each microscope type are outlined.
DARK FIELD MICROSCOPY by SIVASANGARI SHANMUGAM
Dark-field microscopy is ideally used to illuminate unstained samples causing them to appear brightly lit against a dark background.
This type of microscope contains a special condenser that scatters light and causes it to reflect off the specimen at an angle
Bright field microscopy, Principle and applicationsKAUSHAL SAHU
Introduction
History
Basic Component of Microscope
Light Microscopy
Types of Light Microscopy
What Are Bright Microscopy
Principle of Bright Microscope
Advantage
Disadvantage
Application
Conclusion
Reference
BRIGHT FIELD MICROSCOPY by SIVASANGARI SHANMUGAM
bRIGHT FIELD MICROSCOPY is also called a compound microscope. The name bright - field is derived from the fact that the specimen is dark and contrasted by the surrounding bright viewing field.
Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye (objects that are not within the resolution range of the normal eye). There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy.
LIGHT MICROSCOPY by SIVASANGARI SHANMUGAM
The optical microscope, The functions of a light microscope is based on its ability to focus a beam of light through, which is very small and transparent, to produce an image.
during this ppt of microscopes we will be able to know
INTRODUCTION
DEFINITION
HISTORICAL BACKGROUND
VARIABLES USED IN MICROSCOPY
VARIOUS TYPES OF MICROSCOPES
COMPOUND MICROSCOPE - Structure and Function
USE OF MICROSCOPE
CARE OF MICROSCOPE
defintion
A microscope (Greek: micron = small and scopos = aim)
MICROSCOPE - An instrument for viewing objects that are too small to be seen by the naked or unaided eye
MICROSCOPY - The science of investigating small objects using such an instrument is called microscopy
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.
DARK FIELD MICROSCOPY by SIVASANGARI SHANMUGAM
Dark-field microscopy is ideally used to illuminate unstained samples causing them to appear brightly lit against a dark background.
This type of microscope contains a special condenser that scatters light and causes it to reflect off the specimen at an angle
Bright field microscopy, Principle and applicationsKAUSHAL SAHU
Introduction
History
Basic Component of Microscope
Light Microscopy
Types of Light Microscopy
What Are Bright Microscopy
Principle of Bright Microscope
Advantage
Disadvantage
Application
Conclusion
Reference
BRIGHT FIELD MICROSCOPY by SIVASANGARI SHANMUGAM
bRIGHT FIELD MICROSCOPY is also called a compound microscope. The name bright - field is derived from the fact that the specimen is dark and contrasted by the surrounding bright viewing field.
Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye (objects that are not within the resolution range of the normal eye). There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy.
LIGHT MICROSCOPY by SIVASANGARI SHANMUGAM
The optical microscope, The functions of a light microscope is based on its ability to focus a beam of light through, which is very small and transparent, to produce an image.
during this ppt of microscopes we will be able to know
INTRODUCTION
DEFINITION
HISTORICAL BACKGROUND
VARIABLES USED IN MICROSCOPY
VARIOUS TYPES OF MICROSCOPES
COMPOUND MICROSCOPE - Structure and Function
USE OF MICROSCOPE
CARE OF MICROSCOPE
defintion
A microscope (Greek: micron = small and scopos = aim)
MICROSCOPE - An instrument for viewing objects that are too small to be seen by the naked or unaided eye
MICROSCOPY - The science of investigating small objects using such an instrument is called microscopy
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.
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A microscope is an instrument used to see objects that are too small for the naked eye. The science of investigating small objects using such an instrument is called microscopy. Microscopic means invisible to the eye unless aided by a microscop
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The microscope has evolved a lot from the time of Leeuwenhoek. This presentation gives a brief overview about the types of microscope their principle of function and application.
Learn how to make invisible ink and then write with it that can't be seen with naked eye. Learn how to see this invisible writing when you. This is an amazing science experiment. Finally, learn how this works
Presentation to the Gulliver Forum in February 2007 on how my public library is using a blog and how other libraries are useing Web 2.0 tools to provide service to their users.
Microscopy is the technique of using microscopes to observe and analyze objects that are too small to be seen by the naked eye. Microscopes are instruments that magnify and resolve the details of objects, allowing scientists and researchers to study the structure, composition, and behavior of materials and specimens at a microscopic level
Microscopy is the technique of using microscopes to observe and analyze objects that are too small to be seen by the naked eye. Microscopes are instruments that magnify and resolve the details of objects, allowing scientists and researchers to study the structure, composition, and behavior of materials and specimens at a microscopic level
Introduction to microscopy
Different parts of a microscope & their function
Different types of microscopy
Different types of optical microscopy
Different types of electron microscopy
Different terms used in microscopy
Staining- Simple, Differential, Special
Gram Staining
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
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 .
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.
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.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
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.
The ASGCT Annual Meeting was packed with exciting progress in the field advan...
Types of microscope
1.
2. Light Microscope : use sunlight or artificial
light.
1. Bright field microscope.
2. Dark field microscope.
3. Phase contrast microscope.
4. Fluorescence microscope.
Electron microscope : use of electron.
1. Transmission electron microscope.
2. Scanning electron microscope.
3.
4. In light microscopy, light typically passes through a
specimen and then through a series of magnifying
lenses.
5. Microscopes are of great importance in the study of
microorganisms and biomolecules.
Light microscopes are simplest of all microscopes.
Light microscopes use lenses to bend and focus light
rays to produce enlarged images of small objects.
7. Light is transmitted and focussed by mirror and
condenser.
Focussed light illuminate the object or specimen.
The refracted light is collected by an objective
where primary image of the object is formed, it is
real,inverted enlarged image of the object.
The eyepiece further magnifies this primary
image into virtual,erect enlarged image, this is
the final image that lies above the stage.
8. Total magnification of specimen by multiplying the
objective lens magnification power by the ocular lens
magnification power.
Low power x 10, high power x 40 and oil immersion x
100
9. Is also called resolving power of image.
i.e the ability to distinguish that two objects are
separate and not one.
Resolving power of a microscope is determined by
the wave length of light entering the objective lens.
A general principle of ,microscopy is that the shorter
the wave length of light used in the instrument, the
greater the resolution
10. The white light used in a compound light
microscope has relatively long wave length and
cannot resolve structures smaller than about 0.2
µm.
Immersion oil is placed between the glass and
objective lens.
The immersion oil has the same refractive index
as glass of the microscope.
The oil enhances the resolution by preventing
light rays from dispersing and changing wave
length after passing through the specimens.
11. Observation of morphology of microorganisms.
Detection of cell structures.
Observation of intracellular structures.
Observation of motility.
Measurement of size.
Observation of blood smears.
12.
13.
14. The useful magnification of Light microscope is
limited by its resolving power.
The resolving power in limited by wavelength of
illuminating beam.
Resolution is determine by certain physical
parameters like wave length of light and light
generating power of the objective & condenser lens.
15. The ordinary microscope is called as a brightThe ordinary microscope is called as a bright
field microscop.field microscop.
It forms dark image against bright background.It forms dark image against bright background.
16.
17.
18.
19. Image is created by objective and ocular lenses
working together.
Light from illuminated specimen is focused by the
objective lens creating enlarged image within the
microscope.
The ocular lens further modifies the primary image.
20. Total magnification is calculated by magnification by
objective multiply by magnification by eyepiece.
Ex : 45x X 10x =450x
21.
22. Bright field compound microscopes are commonly
used to view live and immobile specimens such as
bacteria, cells, and tissues. For transparent or
colorless specimens, however, it is important that
they be stained first so that they can be properly
viewed under this type of a microscope. Staining is
achieved with the use of a chemical dye. By applying
it, the specimen would be able to adapt the color of
the dye. Therefore, the light won’t simply pass
through the body of the specimen showing nothing
on the microscope’s view field
23.
24. Dark field microscopy is frequently performed on the
same microscope on which bright-field microscopy is
performed.
Instead of the normal condenser that contains an
opaque disk.
The disk blocks light that would enter the objective
lens directly.
25. Only light that has been reflected or refracted by the
specimen forms the image
The field surrounding specimen appears dark while
the object brightly illuminated
26.
27. The advantage of darkfield microscopy also becomes
its disadvantage: not only the specimen, but dust and
other particles scatter the light and are easily
observed
For example, not only the cheek cells but the bacteria
in saliva are evident.
The dark field microscopes divert illumination and
light rays thus, making the details of the specimen
appear luminous.
28. Dark field light microscopes provide good results,
especially through the examination of live blood
samples.
It can yield high magnifications of living bacteria and
low magnifications of the tissues and cells of certain
organisms.
Certain bacteria and fungi can be studied with the
use of dark field microscopes.
29.
30. The principle of phase-contrast microscopy is basedThe principle of phase-contrast microscopy is based
on the wave nature of light rays and the fact thaton the wave nature of light rays and the fact that
light rays can be in phase or out of phase.light rays can be in phase or out of phase.
In a phase-constrast microscope, one set of light raysIn a phase-constrast microscope, one set of light rays
comes directly from the light sources.comes directly from the light sources.
The other set comes from light that is reflected orThe other set comes from light that is reflected or
diffracted from a particular structure in thediffracted from a particular structure in the
specimen.specimen.
(diffraction is the scattering of light rays-direct rays(diffraction is the scattering of light rays-direct rays
and reflected or diffracted rays are broughtand reflected or diffracted rays are brought
together)together)
31. Both combined rays form an image of the specimen
on the ocular lens, containing areas that are relatively
light (in-phase), through shades of gray, to black(out
phase)
Through the use of annular lens, the differences in
phase are amplified so that in-phase light appears
brighter than out-of-phase.
32.
33. To study unstained livingcells.To study unstained livingcells.
Detailed examination of internal structures inDetailed examination of internal structures in
living microorganismliving microorganism
To study flagellar movements and motility ofTo study flagellar movements and motility of
bacteria and protozoans.bacteria and protozoans.
To study intestinal and other living protozoaTo study intestinal and other living protozoa
such as amoeba and trichomonas.such as amoeba and trichomonas.
To examine fungi grown in cultureTo examine fungi grown in culture
34. Phase contrast microscopy is used in study of living
cells and tissues.
Microbes and parasites can be study .
Useful in observing cells cultured in vitro during
mitosis.
35.
36. In all types of microscopes, cell constituents are not
distinguishable, although staining dose , but not
totally.
In fluorescent microscopy, various fluorescent dyes
are used which gives property of fluorescence to only
specific part of the cell and hence it can be focused.
37. When certain compounds are illuminated with high
energy light, they then emit light of a different, lower
frequency. This effect is known as fluorescence.
Often specimens show their own characteristic
autofluorescence image, based on their chemical
makeup.
38. Many different fluorescent dyes can be used to
stain different structures or chemical
compounds.
One particularly powerful method is the
combination of antibodies coupled to a
fluorochrome as in immunostaining.
Examples of commonly used fluorochromes are
fluorescein or rhodamine
39.
40. A component of interest in the specimen isA component of interest in the specimen is
specifically labeled with a fluorescent moleculespecifically labeled with a fluorescent molecule
called a fluorophorecalled a fluorophore
The specimen is illuminated with light of a specificThe specimen is illuminated with light of a specific
wavelength (or wavelengths) which is absorbed bywavelength (or wavelengths) which is absorbed by
the fluorophores, causing them to emit longerthe fluorophores, causing them to emit longer
wavelengths of light (of a different color than thewavelengths of light (of a different color than the
absorbed light).absorbed light).
Typical components of a fluorescence microscopeTypical components of a fluorescence microscope
are the light source (xenon arc lamp or mercury-are the light source (xenon arc lamp or mercury-
vapor lamp), the excitation filter, the dichroicvapor lamp), the excitation filter, the dichroic
mirror and the emission filter.mirror and the emission filter.
41. The illumination light is separated from theThe illumination light is separated from the
much weaker emitted fluorescence through themuch weaker emitted fluorescence through the
use of an emission filter.use of an emission filter.
The filters and the dichroic are chosen to matchThe filters and the dichroic are chosen to match
the spectral excitation and emissionthe spectral excitation and emission
characteristics of the fluorophore used to labelcharacteristics of the fluorophore used to label
the specimen.the specimen.
In this manner, a single fluorophore (color) isIn this manner, a single fluorophore (color) is
imaged at a time. Multi-color images of severalimaged at a time. Multi-color images of several
fluorophores must be composed by combiningfluorophores must be composed by combining
several single-color images.several single-color images.
42.
43.
44.
45.
46. Fluorescence microscopy is a critical tool for
academic and pharmaceutical research, pathology,
and clinical medicine.
47.
48. Electron microscopy is in some ways
comparable to light microscopy. Rather
than using glass lenses, visible light, and
the eye to observe the specimen, the
electron microscope uses electromagnetic
lenses, electrons, and a fluorescent screen
to produce the magnified image. That
image can be captured on photographic
film to create an electron photomicrograph.
49. The superior resolution of the electron
microscope is due to the fact that electrons have
a much shorter wavelenght than the photons of
white light.
The electron beam is focused by circular electron
magnets, which are analogous to the lenses in
the light microscope. The object which is held in
the path of the beam scatters the electrons and
produces an image which is focused on a
fluorescent viewing screen.
50.
51. A. Transmission Electron Microscope
B. Scanning Electron Microscope
A. Transmission Electron Microscope
In a transmission microscope, electrons pass through
the specimen and scattered. Magnetic lenses focus
the image onto a fluorescent screen or photographic
plate.
Electron light pass directly through the specimen
that has been prepared by thin sectioning, freeze
fracturing, or freeze etching.
It is used to observe fine details of cell structure.
52.
53. B. Scanning Electron Microscope
In a scanning electron microscope, primary
electron sweep across the specimen and knock
electrons from its surface.
The second electron is picked up by a collector,
amplified, and transmitted onto viewing screen
or photographic plate.
It scan a beam of electrons back and forth over
the surface of a specimen producing three-
dimensional views of the surfaces of whole
microorganism.