The document provides an overview of microscopy including:
1. It discusses the historical development of the microscope from its invention in the 1590s to improvements made by scientists like Hooke, Huygens, and van Leeuwenhoek in the 1600s-1700s.
2. It describes several types of microscopes like brightfield, darkfield, phase contrast, fluorescence, electron (TEM and SEM), confocal, and scanning probe microscopes; and explains their basic optical principles and uses.
3. It covers various optical components and concepts in microscopy like magnification, resolution, numerical aperture, aberration, contrast, and different techniques like micrometry.
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.
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.
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.
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.
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.
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
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
Microscopy - Magnification, Resolving power, Principles, Types and ApplicationsNethravathi Siri
Magnification, Resolving power, Principles and Applications of Simple, Compound, Stereozoom, Phase contrast, Fluorescent and Electron microscopes (TEM & SEM).
Microscopy is the technical field that uses microscopes to observe samples which are not in the resolution range of the normal-unaided eye.
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
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.
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.
MICROMETRY is a technique used to measure the size of microscopic objects.It is based on principle of Calibration of the ocular micrometer using the stage micrometer. If you change microscope, the calibration process must be done again for each of the objective lenses that you are using because the magnification is different on different microscopes.
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.
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
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
Microscopy - Magnification, Resolving power, Principles, Types and ApplicationsNethravathi Siri
Magnification, Resolving power, Principles and Applications of Simple, Compound, Stereozoom, Phase contrast, Fluorescent and Electron microscopes (TEM & SEM).
Microscopy is the technical field that uses microscopes to observe samples which are not in the resolution range of the normal-unaided eye.
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
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.
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.
MICROMETRY is a technique used to measure the size of microscopic objects.It is based on principle of Calibration of the ocular micrometer using the stage micrometer. If you change microscope, the calibration process must be done again for each of the objective lenses that you are using because the magnification is different on different microscopes.
This presentation is about the introduction of microscopy, its history, parts of a microscope and different types of microscopes along with a brief discussion of their working principles.
A **bright field microscope** is a type of compound light microscope that illuminates the background against a stained specimen ¹². It is commonly used in practical labs to study organisms' behavior and characteristics such as size, shape, and arrangement ². The microscope uses light rays to produce a dark image against a bright background ¹. It is specially designed with magnifying glasses known as lenses that modify the specimen to produce an image seen through the eyepiece ¹. The bright field microscope is made up of various parts, including the eyepiece, objective lenses, focusing knobs, and stage ¹.
I hope this helps!
Source: Conversation with Bing, 7/11/2023
(1) Brightfield Microscope (Compound Light Microscope)- Definition .... https://microbenotes.com/brightfield-microscope/.
(2) Bright Field Microscopy - Biology Reader. https://biologyreader.com/bright-field-microscopy.html.
(3) Bright-field microscopy - Wikipedia. https://en.wikipedia.org/wiki/Bright-field_microscopy.
(4) Bright Field Microscope: Definition, Parts, Working Principle, Application. https://microbiologynote.com/bright-field-microscope-definition-parts-working-principle-application/.
these slides gives information about the types of microscopy. microscopy is divided into two type on the bases of their application which are stereoscopic and biological microscope. another types of microscopy are optical microscope, electron microscope, x-ray microscope and also scanning probe microscopy which perform many function.
Research is "creative and systematic work undertaken to increase the stock of knowledge". It involves the collection, organization and analysis of evidence to increase understanding of a topic, characterized by a particular attentiveness to controlling sources of bias and error.
Microscope and Microscopy
Principal , Function & Difference of various types of Light & Electron microscope.Microscopy is the technical field of using microscopes to view samples & objects that cannot be seen with the unaided eye (objects that are not within the resolution range of the normal eye).
Microscopists explore the relationships between structures & properties for a very wide variety of materials ranging from soft to very hard, from inanimate materials to living organisms, in order to better understand it. Zachariaz Janssen 1585 Robert Hooks 1665
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2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
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2. LAYOUT OF PRESENTATION
• Introduction
• Historical Background
• Variables Used In Microscopy
• Parts of microscope
• Micrometry
• Types of Microscope & Uses
• Care Of Microscope
3.
4. - A microscope (Greek: mikron = small and
scopeos = to look).
- MICROSCOPE: Is 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.
5.
6. • 1590 - Hans Janssen and his son Zacharias
Janssen, developed first microscope.
• 1609 - Galileo Galilei - occhiolino or compound
microscope.
• 1620 - Christian Huygens, another Dutchman,
developed a simple 2-lens ocular system that was
chromatically corrected.
9. 1.MAGNIFICATION
• Degree of enlargement.
• No of times the length, breadth or diameter, of
an object is multiplied.
• It depends upon – Optical tube length
Focal length of objective
Magnifying power of eye piece
• TOTAL MAGNIFICATION:
magnification of the eyepiece x magnification of
the objective.
10. 2.RESOLUTION
• Ability to reveal closely adjacent structural
details as separate and distinct.
• LIMIT OF RESOLUTION (LR): The minimum
distance between two visible bodies at which
they can be seen as separate and not in contact
with each other.
• LR = 0.61 x W W = Wavelength
NA NA = Numerical aperature
11. 3.NUMERICALAPERTURE(NA)
- Ratio of diameter of lens to its focal length
- NA = n Sin θ/2
n = refractive index,
θ = angle of aperture (CAD)
θ/2
A
B DC
4.DEFINITION
Capacity of an objective to render outline of
the image of an object clear and distinct.
20. • Helps in measurement of the size of
microscopic objects.
• Achieved by the use of-
1.STAGE MICRO METER
2.MICOMETER EYE PIECE
21. CALIBRATION OF THE
OCULAR MICROMETER
Stage micrometer
(objective micrometer)
has calibration lines or
graduations that are
separated 0.01 mm (10
µm) apart.
Calibration achieved by
superimposing the two
scales and determining
how many ocular
graduation coincide with
graduations on stage
micrometer.
Ocular micrometer with retaining ring inserted into
the eyepiece’s base
Stage micrometer positioning by centering the
small glass disk over the light source
After completed calibration, specimen’s slide is
positioned for measurement
22.
23.
24. LIGHT MICROSCOPE : use sunlight or
artificial light.
A. Bright field microscope.
B. Dark field microscope.
C. Phase contrast microscope.
D. Fluorescence microscope.
ELECTRON MICROSCOPE : use of
electron.
1. Transmission electron microscope.
2. Scanning electron microscope.
25. A. BRIGHT-FIELD MICROSCOPE
Produces a dark image against a brighter
background.
Has several objective lenses.
2 types: Simple
Compound
26. SIMPLE
• Contain a single magnifying lens.
COMPOUND
• Series of lenses for magnification.
• Light passes through specimen into objective lens
• Oil immersion lens increases resolution.
• Have one or two ocular lenses.
• Resolution=200nm
29. ADVANTAGES
• Used to view live or stained cells.
• Simple setup with very little preparation required.
DISADVANTAGES
• Biological specimen are often of low contrast and
need to be stained.
• Staining may destroy or introduce artifacts.
• Resolution is limited to 200nm.
31. B.DARK FIELD MICROSCOPE
• Produces a bright image of the object against a dark
back ground.
• Optical system to enhance the contrast of unstained
bodies.
• Specimen appears gleaming bright against dark
background.
32. 32
REQUISITES FOR DARK FIELD
MICROSCOPY
- Dark ground
condenser
- High intensity lamp
- Funnel stop
34. USES OF DARK GROUND
MICROSCOPY
Treponema pallidum
Useful in demonstrating
-Treponema pallidum
- Leptospira
- Campylobacter jejuni
- Endospore
35. ADVANTAGES
• Simple setup
• Provides contrast to unstained tissue,so
living cells can be viewed.
DISADVANTAGES
• Specimen needs to be strongly illuminated
which can damage delicate samples.
36. PHASE CONTRAST MICROSCOPE
• First described in 1934 by Dutch physicist Frits
Zernike
• Produces high-contrast images of transparent
specimens.
• Advantage - Living cells can be examined in their
natural state.
37. PRINCIPLE OF PHASE CONTRAST
MICROSCOPY
• It is an optical illumination technique in which small phase
shifts in the light passing through a transparent specimen
are converted into contrast changes in the image.
• Light rays in phase produce brighter image.
• Light rays out of phase form darker image.
• Contrast is due to out of phase rays.
40. CONDENSER ANNULUS
• The condenser annulus or annular diaphragm is
opaque flat-black (light absorbing) plate with a
transparent annular ring.
• Produces hollow cone of light.
41. PHASE PLATE
- Placed in back focal plane of objective.
- Function:
1. Enhances phase difference by retarding
diffracted wave front by one quarter of
wavelength .
2. Reduces intensity of direct rays and
equalizes it with diffracted rays intensity.
44. ADVANTAGES
• Phase contrast enables visualization of internal
cellular components.
• Diagnosis of tumor cells .
• Examination of growth, dynamics, and behavior
of a wide variety of living cells in cell culture.
• Ideal for studying & interupting thin specimen.
45. DISADVANTAGES
• Annuli or ring limits the apperature to some
extents which causes decrease in resolution.
• Not ideal for thick specimen.
• Shade off and Halo effect may occur.
46. THE FLUORESCENCE MICROSCOPE
• Exposes specimen to ultraviolet, violet, or blue
light.
• Specimens usually stained with fluorochromes.
• Shows a bright image of the object resulting
from the fluorescent light emitted by the
specimen.
47. PRINCIPLE OF FLUORESCENCE
MICROSCOPY
• Certain dyes,called as fluorochrome after
absorbing UV rays raised to a higher energy
level.
• When the dye molecules return to their normal
state,they release excess energy in the form of
visible light(fluoroscence).
49. 49
USE OF FLUORESCENCE
MICROSCOPY
- Auramine Rhodamine – Yellow
fluorescence Tubercle bacilli
- Acridine Orange R - gives orange
red fluorescence with RNA and
yellow green fluorescence with
DNA
- QBC
- IMMUNOFLUORESCENCE
51. 51
ELECTRON
MICROSCOPE
Co-invented by Max knoll
and Ernst Ruska in 1931.
Electron Microscopes uses
a beam of highly energetic
electrons to examine
objects on a very fine
scale.
Magnification can upto
2million times while best
light microscope can
magnify up to 2000 times.
52. TRANSMISSION ELECTRON
MICROSCOPE (TEM)
Stream of electrons is formed.
Accelerated using a positive electrical potential.
Focused by metallic aperture and Electro
magnets.
Interactions occur inside the irradiated sample
which are detected and transformed into an
image.
55. - Scan a gold-plated specimen to give a 3-D view
of the surface of an object which is black and
white.
- Used to study surface features of cells and
viruses.
- Scanning Electron microscope has resolution
1000 times better than Light microscope.
SCANNING ELECTRON
MICROSCOPE
58. COMPARING SEM AND TEM
TEM SEM
Imaging
Electrons must pass through and
be transmitted by the specimen
Information needed is
collected near the surface
of the specimen
Electron Beam Broad, static beams
Beam focused to fine point;
sample is scanned line by line
Voltages Needed
TEM voltage ranges from
60-300,000 volts
Accelerating voltage much lower; not
necessary to penetrate the specimen
Image Rendering
Transmitted electrons are
collectively focused by the
objective lens and magnified to
create a real image
Beam is scanned along the surface of
the sample to
build up the image
Interaction of the
beam electrons
Specimen must be very thin
Wide range of specimens allowed;
simplifies
sample preparation
59.
60. Uses a laser beam to illuminate a
specimen whose image is then
digitally enhanced for viewing on a
computer monitor.
Laser beam scans single plane of
1µm thickness.
CONFOCAL LASER SCANNING
MICROSCOPE
63. USES OF CONFOCAL MICROSCOPE
- Observing cellular morphology in multilayered
specimen.
- Eg. used in diagnosing Ca cervix
- Evaluation and diagnosis of basal cell carcinoma
of skin.
64. ADVANTAGE OF USING A
CONFOCAL MICROSCOPE
- By having a confocal pinhole, the microscope is
really efficient at rejecting out of focus
fluorescent light so that very thin section of a
sample can be analyzed.
- By scanning many thin sections through a
sample, one can build up a very clean three-
dimensional image .
65. INVERTED MICROSCOPE
• Used in metallurgy
• Examination of cultures in flat bottom dishes
• Micro dissection
• Examination of parasites
• Observation of agglutination in serology
66. STEREO MICROSCOPE
• Double Microscope
• Produces 3D images
POLARIZING MICROSCOPE
• Uses two Polariser
• Gives information about Birefringence of a body
• Used in Crystallography, Urine examination
• Apple Green Birefringerence in AMYLODOSIS
67. SCANNING PROBE MICROSCOPE
- Class of Microscope that measures surface
features by moving a sharp probe over object
surface. Used to visualize atoms and molecules
• scanning tunneling microscope (stm)
• atomic force microscope (afm)
68. SCANNING
TUNNELING
MICROSCOPE
Steady current
(tunneling current)
maintained between
microscope probe and
specimen.
The arrangement of
atoms on the specimen
is determined by moving
probe tip back and froth
over specimen keeping a
constant height.
ATOMIC FORCE
MICROSCOPE
Sharp probe moves
over surface of
specimen at constant
distance.
Up and down
movement of probe as
it maintains constant
distance is detected
and used to create
image.
69. SCANNING PROBE MICROSCOPES
Scanning probe microscopes
“feel” changes in surfaces of
the sample. They use ultra-
sharp tips to measure
changes in electric currents,
electrostatic forces, or
magnetic forces coming from
the surface.
The tips of the microscopes have to
be incredibly sharp to allow them
to collect information about
individual atoms or molecules.
70.
71. • PROPER STORAGE
• HANDLING
• CARE OF LENSES
• CARE OF OIL EMERSION OBJECTIVE
• CARE OF LAMP