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Introduction to the
Microscope
• History
• Types
• Care
• Parts & functions
• Focusing
History of the Microscope
• 1590 –first compound
microscope
History of the Microscope
• 1655 – Robert Hooke used a
compound microscope to
observe pores in cork
–He called them “cells”
History of the Microscope
• Antoine van Leeuwenhoek
–1st to see single-celled
organisms in pond water
Microscope Vocabulary
• Magnification: increase of an
object’s apparent size
• Resolution: power to show
details clearly
• Both are needed to see a clear
image
Types of Microscopes
• 1. Compound Light Microscope
–1st type of microscope, most
widely used
–light passes through 2 lenses
–Can magnify up to 2000x
Ocular lens
Objective
lenses
Types of Microscopes
• 2. Electron Microscope
–Used to observe VERY small
objects: viruses, DNA, parts of
cells
–Uses beams of electrons rather
than light
–Much more powerful
Types of Microscopes
• Transmission Electron
Microscope (TEM)
–Can magnify up to 250,000x
Types of Microscopes
• Scanning Electron Microscope
(SEM)
–Can magnify up to 100,000x
Circa 1000AD –
The first vision aid
was invented
(inventor unknown)
called a reading
stone. It was a glass sphere that
magnified when laid on top of reading
materials.
Circa 1284 –
Italian, Salvino D'Armate is credited
with inventing the first wearable eye
glasses.
1590 – Two Dutch eye glass makers,
Zaccharias Janssen and son Hans Janssen
experimented with multiple lenses
placed in a tube. The Janssens observed
that viewed objects in front of the tube
appeared greatly enlarged, creating both
the forerunner of the compound
microscope and the telescope.
1665 – English
physicist, Robert
Hooke looked at a
sliver of cork
through a
microscope lens and
noticed some
"pores" or "cells" in
it.
1674 – Anton van Leeuwenhoek built a
simple microscope with only one lens to
examine blood, yeast, insects and many
other tiny objects. Leeuwenhoek was the
first person to describe bacteria, and he
invented new methods for grinding and
polishing microscope lenses that allowed
for curvatures providing magnifications of
up to 270 diameters, the best available
lenses at that time.
18th century – Technical innovations
improved microscopes, leading to
microscopy becoming popular among
scientists. Lenses combining two types
of glass reduced the "chromatic effect"
the disturbing halos resulting from
differences in refraction of light.
1830 – Joseph Jackson Lister reduces
spherical aberration or the "chromatic
effect" by showing that several weak
lenses used together at certain distances
gave good magnification without
blurring the image. This was the
prototype for the compound microscope.
1872 – Ernst Abbe, then research
director of the Zeiss Optical Works,
wrote a mathematical formula called the
"Abbe Sine Condition". His formula
provided calculations that allowed for
the maximum resolution in microscopes
possible.
1903 – Richard
Zsigmondy developed
the ultramicroscope
that could study
objects below the
wavelength of light.
He won the Nobel
Prize in Chemistry in
1925.
1932 – Frits Zernike
invented the phase-
contrast microscope
that allowed for the
study of colorless and
transparent biological
materials for which he
won the Nobel Prize
in Physics in 1953.
1931 – Ernst Ruska co-invented the
electron microscope for which he won the
Nobel Prize in Physics in 1986. An
electron microscope depends on electrons
rather than light to view an object,
electrons are speeded up in a vacuum
until their wavelength is extremely short,
only one hundred-thousandth that of
white light. Electron microscopes make
cockroach antenna
pigeon blood
1931 – Ernst
Ruska
it possible to
view objects
as small as
the diameter
of an atom.
1981 – Gerd Binnig and Heinrich Rohrer
invented the scanning tunneling
microscope that gives three-dimensional
images of objects down to the atomic
level. Binnig and Rohrer won the Nobel
Prize in Physics in 1986. The powerful
scanning tunneling microscope is the
strongest microscope to date.
•Compound Microscope
•Dissection Microscope
•Scanning Electron Microscope (SEM)
•Transmission Electron Microscope (TEM)
Compound microscopes are light
illuminated. The image seen with this type
of microscope is two dimensional. This
microscope is the most commonly used.
You can view individual cells, even living
ones. It has high magnification. However,
it has a low resolution.
Frog’s blood
1,000x
Paulownia Wood c.s.
200x
A dissection microscope is light
illuminated. The image that appears is
three dimensional. It is used for dissection
to get a better look at the
larger specimen. You cannot
see individual cells because
it has a low magnification.
(also called stereo
microscope)
Sunflower with moth
pupa in the stem
10x
Head of a moth pupa
60x
SEM use electron illumination. The
image is seen in 3-D. It has high
magnification and high resolution. The
specimen is coated in gold
and the electrons bounce
off to give you and exterior
view of the specimen. The
pictures are in black and
white.
TEM is electron illuminated. This gives a
2-D view. Thin slices of specimen are
obtained. The electron beams pass
through this. It has
high magnification
and high resolution.
mitochondrion
bacillus bacteria
dividing
• Always carry with 2 hands
• Never touch the lenses with your fingers.
• Only use lens paper for cleaning
• Do not force knobs
• Keep objects clear of desk and cords
• When you are finished with your "scope",
rotate the nosepiece so that it's on the low
power objective, roll the stage down to
lowest level, rubber band the cord, then
replace the dust cover.
Ocular lens
Body Tube
Revolving Nosepiece
Arm
Objective Lens
Stage
Stage
Clips Coarse adjustment knob
Fine adjustment knob
Base
Diaphragm
Light
Ocular lens
magnifies; where you
look through to see the
image of your specimen.
They are usually 10X or
15X power. Our
microscopes have an ocular
lens power of 10x.
arm
supports the tube and
connects it to the
base
stage
the flat platform
where you place
your slides
coarse adjustment knob
moves stage (or body
tube) up and down
fine adjustment knob
small, round knob on
the side of the
microscope used to
fine-tune the focus of
your specimen
after using the coarse
adjustment knob
base
the bottom of the
microscope, used for
support
body tube
connects the eyepiece
to the objective
lenses
revolving nosepiece
the part that holds two
or more objective lenses
and can be rotated to
easily change power
objective lens
Adds to the magnification
Usually you will find 3 or
4 objective lenses on a
microscope. They almost
always consist of 4X, 10X,
40X and 100X
powers. When coupled
with a 10X (most common)
objective lenses
eyepiece lens, we get total
magnifications of 40X (4X
times 10X), 100X , 400X
and 1000X.
The shortest
lens is the lowest power, the
longest one is the lens with
the greatest power. Lenses
are color coded.
objective lenses
The high power objective
lenses are retractable (i.e.
40XR). This means that if
they hit a slide, the end of
the lens will
push in (spring loaded)
thereby protecting the lens
and the slide.
stage clips
Stage clips hold the slides in
place. If your microscope
has a mechanical stage, you
will be able to move the
slide around by turning two
knobs. One
moves it left and right, the
other moves it up and down.
diaphragm
controls the amount of light
going through the specimen
Many microscopes have a
rotating disk under the
stage. This diaphragm has
different sized holes and is
used to vary the intensity
and size of the cone of light
diaphragm
that is projected upward into
the slide. There is no set rule
regarding which setting to use
for a particular power. Rather,
the setting is a function of the
transparency of the specimen,
the degree of contrast
you desire and the particular
objective lens in use.
light
makes the specimen
easier to see
The proper way to focus a microscope is
to start with the lowest power objective
lens first and while looking from the
side, crank the lens down as close to the
specimen as possible without touching
it. Now, look through the eyepiece lens
and focus upward only until the image
is sharp. If you can't get it in focus,
repeat the process again.
Once the image is sharp with the low
power lens, you should be able to
simply click in the next power lens and
do minor adjustments with the focus
knob. If your microscope has a fine
focus adjustment, turning it a bit should
be all that's necessary. Continue with
subsequent objective lenses and fine
focus each time.
Rotate to 40x objective, locate desired
portion of specimen in the center of the
field. Refocus very carefully so that the
specimen is focused as sharply as
possible. (Do not
alter focus for the
Following steps )
Partially rotate so that 40x and 100x
objectives straddle the specimen.
Place a small drop of oil on the slide in
the center of the lighted area. (Take care
not to dribble on the stage.)
Put the small drop
of oil directly over
the area of the
specimen to be
Examined.
Rotate so that the 100x oil immersion
objective touches the oil and clicks into
place.
Focus only with fine focus. Hopefully,
the specimen will come into focus easily.
Do not change focus dramatically.
Clean up!: When you have finished for
the day, wipe the 100x oil immersion
objective carefully with lens paper to
remove all oil. Wipe oil from the slide
thoroughly with a Kimwipe. Cleanse
stage should any oil have spilled on it.
Recap the immersion oil container
securely, replace in drawer.

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Microscopy use this.ppt

  • 1. Introduction to the Microscope • History • Types • Care • Parts & functions • Focusing
  • 2. History of the Microscope • 1590 –first compound microscope
  • 3. History of the Microscope • 1655 – Robert Hooke used a compound microscope to observe pores in cork –He called them “cells”
  • 4. History of the Microscope • Antoine van Leeuwenhoek –1st to see single-celled organisms in pond water
  • 5. Microscope Vocabulary • Magnification: increase of an object’s apparent size • Resolution: power to show details clearly • Both are needed to see a clear image
  • 6. Types of Microscopes • 1. Compound Light Microscope –1st type of microscope, most widely used –light passes through 2 lenses –Can magnify up to 2000x
  • 8. Types of Microscopes • 2. Electron Microscope –Used to observe VERY small objects: viruses, DNA, parts of cells –Uses beams of electrons rather than light –Much more powerful
  • 9. Types of Microscopes • Transmission Electron Microscope (TEM) –Can magnify up to 250,000x
  • 10.
  • 11. Types of Microscopes • Scanning Electron Microscope (SEM) –Can magnify up to 100,000x
  • 12.
  • 13.
  • 14. Circa 1000AD – The first vision aid was invented (inventor unknown) called a reading stone. It was a glass sphere that magnified when laid on top of reading materials.
  • 15. Circa 1284 – Italian, Salvino D'Armate is credited with inventing the first wearable eye glasses.
  • 16. 1590 – Two Dutch eye glass makers, Zaccharias Janssen and son Hans Janssen experimented with multiple lenses placed in a tube. The Janssens observed that viewed objects in front of the tube appeared greatly enlarged, creating both the forerunner of the compound microscope and the telescope.
  • 17. 1665 – English physicist, Robert Hooke looked at a sliver of cork through a microscope lens and noticed some "pores" or "cells" in it.
  • 18. 1674 – Anton van Leeuwenhoek built a simple microscope with only one lens to examine blood, yeast, insects and many other tiny objects. Leeuwenhoek was the first person to describe bacteria, and he invented new methods for grinding and polishing microscope lenses that allowed for curvatures providing magnifications of up to 270 diameters, the best available lenses at that time.
  • 19. 18th century – Technical innovations improved microscopes, leading to microscopy becoming popular among scientists. Lenses combining two types of glass reduced the "chromatic effect" the disturbing halos resulting from differences in refraction of light.
  • 20. 1830 – Joseph Jackson Lister reduces spherical aberration or the "chromatic effect" by showing that several weak lenses used together at certain distances gave good magnification without blurring the image. This was the prototype for the compound microscope.
  • 21. 1872 – Ernst Abbe, then research director of the Zeiss Optical Works, wrote a mathematical formula called the "Abbe Sine Condition". His formula provided calculations that allowed for the maximum resolution in microscopes possible.
  • 22. 1903 – Richard Zsigmondy developed the ultramicroscope that could study objects below the wavelength of light. He won the Nobel Prize in Chemistry in 1925.
  • 23. 1932 – Frits Zernike invented the phase- contrast microscope that allowed for the study of colorless and transparent biological materials for which he won the Nobel Prize in Physics in 1953.
  • 24. 1931 – Ernst Ruska co-invented the electron microscope for which he won the Nobel Prize in Physics in 1986. An electron microscope depends on electrons rather than light to view an object, electrons are speeded up in a vacuum until their wavelength is extremely short, only one hundred-thousandth that of white light. Electron microscopes make
  • 26. 1931 – Ernst Ruska it possible to view objects as small as the diameter of an atom.
  • 27. 1981 – Gerd Binnig and Heinrich Rohrer invented the scanning tunneling microscope that gives three-dimensional images of objects down to the atomic level. Binnig and Rohrer won the Nobel Prize in Physics in 1986. The powerful scanning tunneling microscope is the strongest microscope to date.
  • 28. •Compound Microscope •Dissection Microscope •Scanning Electron Microscope (SEM) •Transmission Electron Microscope (TEM)
  • 29. Compound microscopes are light illuminated. The image seen with this type of microscope is two dimensional. This microscope is the most commonly used. You can view individual cells, even living ones. It has high magnification. However, it has a low resolution.
  • 31. A dissection microscope is light illuminated. The image that appears is three dimensional. It is used for dissection to get a better look at the larger specimen. You cannot see individual cells because it has a low magnification. (also called stereo microscope)
  • 32. Sunflower with moth pupa in the stem 10x Head of a moth pupa 60x
  • 33. SEM use electron illumination. The image is seen in 3-D. It has high magnification and high resolution. The specimen is coated in gold and the electrons bounce off to give you and exterior view of the specimen. The pictures are in black and white.
  • 34. TEM is electron illuminated. This gives a 2-D view. Thin slices of specimen are obtained. The electron beams pass through this. It has high magnification and high resolution.
  • 36. • Always carry with 2 hands • Never touch the lenses with your fingers. • Only use lens paper for cleaning • Do not force knobs • Keep objects clear of desk and cords • When you are finished with your "scope", rotate the nosepiece so that it's on the low power objective, roll the stage down to lowest level, rubber band the cord, then replace the dust cover.
  • 37. Ocular lens Body Tube Revolving Nosepiece Arm Objective Lens Stage Stage Clips Coarse adjustment knob Fine adjustment knob Base Diaphragm Light
  • 38. Ocular lens magnifies; where you look through to see the image of your specimen. They are usually 10X or 15X power. Our microscopes have an ocular lens power of 10x.
  • 39. arm supports the tube and connects it to the base
  • 40. stage the flat platform where you place your slides
  • 41. coarse adjustment knob moves stage (or body tube) up and down
  • 42. fine adjustment knob small, round knob on the side of the microscope used to fine-tune the focus of your specimen after using the coarse adjustment knob
  • 43. base the bottom of the microscope, used for support
  • 44. body tube connects the eyepiece to the objective lenses
  • 45. revolving nosepiece the part that holds two or more objective lenses and can be rotated to easily change power
  • 46. objective lens Adds to the magnification Usually you will find 3 or 4 objective lenses on a microscope. They almost always consist of 4X, 10X, 40X and 100X powers. When coupled with a 10X (most common)
  • 47. objective lenses eyepiece lens, we get total magnifications of 40X (4X times 10X), 100X , 400X and 1000X. The shortest lens is the lowest power, the longest one is the lens with the greatest power. Lenses are color coded.
  • 48. objective lenses The high power objective lenses are retractable (i.e. 40XR). This means that if they hit a slide, the end of the lens will push in (spring loaded) thereby protecting the lens and the slide.
  • 49. stage clips Stage clips hold the slides in place. If your microscope has a mechanical stage, you will be able to move the slide around by turning two knobs. One moves it left and right, the other moves it up and down.
  • 50. diaphragm controls the amount of light going through the specimen Many microscopes have a rotating disk under the stage. This diaphragm has different sized holes and is used to vary the intensity and size of the cone of light
  • 51. diaphragm that is projected upward into the slide. There is no set rule regarding which setting to use for a particular power. Rather, the setting is a function of the transparency of the specimen, the degree of contrast you desire and the particular objective lens in use.
  • 53. The proper way to focus a microscope is to start with the lowest power objective lens first and while looking from the side, crank the lens down as close to the specimen as possible without touching it. Now, look through the eyepiece lens and focus upward only until the image is sharp. If you can't get it in focus, repeat the process again.
  • 54. Once the image is sharp with the low power lens, you should be able to simply click in the next power lens and do minor adjustments with the focus knob. If your microscope has a fine focus adjustment, turning it a bit should be all that's necessary. Continue with subsequent objective lenses and fine focus each time.
  • 55. Rotate to 40x objective, locate desired portion of specimen in the center of the field. Refocus very carefully so that the specimen is focused as sharply as possible. (Do not alter focus for the Following steps )
  • 56. Partially rotate so that 40x and 100x objectives straddle the specimen.
  • 57. Place a small drop of oil on the slide in the center of the lighted area. (Take care not to dribble on the stage.) Put the small drop of oil directly over the area of the specimen to be Examined.
  • 58. Rotate so that the 100x oil immersion objective touches the oil and clicks into place.
  • 59. Focus only with fine focus. Hopefully, the specimen will come into focus easily. Do not change focus dramatically.
  • 60. Clean up!: When you have finished for the day, wipe the 100x oil immersion objective carefully with lens paper to remove all oil. Wipe oil from the slide thoroughly with a Kimwipe. Cleanse stage should any oil have spilled on it. Recap the immersion oil container securely, replace in drawer.