Microscope Chap 5

7/29/2019 Microscope Chap 5

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CHAPTER 5

MICROSCOPE
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-Device that can be used to observe the objects that are toosmall to be seen by the naked instrument or unaided eye.

-The science of investigating small objects using such aninstrument is called microscopy.

-The term microscopic means minute or very small, not visible

with the eye unless aided by a microscope.

-Anton Van Leeuwenhoek's new, improved microscopeallowed people to see things no human had ever seen before.

-The first useful microscope was developed in theNetherlands between 1590 and 1608
http://en.wikipedia.org/wiki/Laboratory_equipmenthttp://en.wikipedia.org/wiki/Eyehttp://en.wikipedia.org/wiki/Sciencehttp://en.wikipedia.org/wiki/Microscopyhttp://en.wikipedia.org/wiki/Microscopichttp://en.wikipedia.org/wiki/Anton_Van_Leeuwenhoekhttp://en.wikipedia.org/wiki/Anton_Van_Leeuwenhoekhttp://en.wikipedia.org/wiki/Anton_Van_Leeuwenhoekhttp://en.wikipedia.org/wiki/Anton_Van_Leeuwenhoekhttp://en.wikipedia.org/wiki/Anton_Van_Leeuwenhoekhttp://en.wikipedia.org/wiki/Anton_Van_Leeuwenhoekhttp://en.wikipedia.org/wiki/Anton_Van_Leeuwenhoekhttp://en.wikipedia.org/wiki/Microscopichttp://en.wikipedia.org/wiki/Microscopyhttp://en.wikipedia.org/wiki/Sciencehttp://en.wikipedia.org/wiki/Eyehttp://en.wikipedia.org/wiki/Laboratory_equipment


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-There are THREE types of microscopes:

- Compound Light Microscope- Electron Microscope Transmission

Electron

Microscope(TEM)

Scanning Electron

Microscope (SEM)


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COMPOUND LIGHT MICROSCOPE

-The term light refers to the method by which lighttransmits the image to your eye. Compounddeals with themicroscope having more than one lens.

-Microscope is the combination of two words; "micro"meaning small and "scope" meaning view.

-The image seen with this type of microscope is two

dimensional.

-This microscope is the most commonly used.


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COMPOUND LIGHT MICROSCOPE


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Diagram Showing Light Traveling Through The

Microscope


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Magnifying Objects/ Focusing Image:

-When viewing a slide through the microscope makesure that the stage is all the way down and the 4Xscanning objective is locked into place.

-Place the slide that you want to view over the apertureand gently move the stage clips over top of the slide tohold it into place.

-Beginning with the 4X objective, looking through theeyepiece making sure to keep both eyes open (if youhave trouble cover one eye with your hand) slowly movethe stage upward using the coarse adjustment knob untilthe image becomes clear.
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-To magnify the image to the next level rotate thenosepiece to the 10X objective. While looking

through the eyepiece focus the image into view usingonly the fine adjustment knob, this should only take a

slight turn of the fine adjustment knob to complete

this task.

-To magnify the image to the next level rotate the

nosepiece to the 40X objective. While looking

through the eyepiece focus the image into view using

only the fine adjustment knob, this should only take aslight turn of the fine adjustment knob to complete

this task.
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TotalMagnification:

-To figure the total magnification of an image that youare viewing through the microscope is really quite

simple. To get the total magnification take the power

of the objective (4X, 10X, 40x) and multiply by the

power of the eyepiece, usually 10X.


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IMAGES FROM COMPOUND LIGHT

MICROSCOPE

Elodea Elodea Elodea

40X 100X 400X


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Microscope Care & Handling

Importance of care

So why do I need to know how to use the microscope?

Because microscopes cost several hundred dollars it is veryimportant to make them last for a long time.

What happens if I break a microscope?

You break it, you buy a new one.....

How long will a microscope last if I take good care of it?

They can last for at least 10 years if you care for the "scope" as well as you care foryour hair
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Care and Handling

Transporting:

When you pick up the microscope and walk with it, grab the arm with one handand place your other hand on the bottom of the base. DON'T SWING THEMICROSCOPE !

Handling & Cleaning:

Never touch the lenses with your fingers. Your body produces an oil that smudges theglass. This oil can even etch the glass if left on too long. Use only LENS PAPER toclean the glass. TOILET PAPER, KLEENEX, AND PAPER TOWELS HAVE FIBERSTHAT CAN SCRATCH THE LENSES.

Storage:

When you are finished with your "scope" assignment, rotate the nosepiece sothat it's on the low power objective, roll the nosepiece so that it's all the waydown to the stage, then replace the dust cover. DON'T FORGET TO USEPROPER TRANSPORTING TECHNIQUES!
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Using the Microscope

Follow these directions when using the microscope!

1. To carry the microscope grasp the microscopes arm with

one hand. Place your other hand under the base.

2. Place the microscope on a table with the arm toward you.

3. Turn the coarse adjustment knob to raise the body tube.

4. Revolve the nosepiece until the low-power objective lens

clicks into place.

5. Adjust the diaphragm. While looking through the

eyepiece, also adjust the mirror until you see a bright whitecircle of light.

6. Place aslide on the stage. Center the specimen over the

opening on the stage. Use the stage clips to hold the slide in

place.
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6. Place aslide on the stage. Center the specimen over the opening on

the stage. Use the stage clips to hold the slide in place.

7. Look at the stage from the side. Carefully turn the coarse adjustment

knob to lower the body tube until the low power objective almost

touches the slide.

8. Looking through the eyepiece, VERY SLOWLY the coarse

adjustment knob until the specimen comes into focus.

9. To switch to the high power objective lens, look at the microscope

from the side. CAREFULLY revolve the nosepiece until the high-

power objective lens clicks into place. Make sure the lens does not hitthe slide.

10. Looking through the eyepiece, turn the fine adjustment knob until

the specimen comes into focus.
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ELECTRON MICROSCOPE

-Electron microscope were developed due to the limitations of

light microscope

-Early 1930s there was a scientific desire to see the fine details ofthe interior structures of organic cells (nucleus, mitochondria..)

-This required 10,000 plus magnification which was just notpossible using Light Microscope

What is Electron Microscopes?

Electron Microscopes are scientific instruments that use a beamof highly energetic electrons to examine objects on a very finescale.


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Electron Microscopes(EMs) function exactly as their optical counterparts except

that they use a focused beam of electrons instead of light to "image" the specimen

and gain information as to its structure and composition.The basic steps involved in all EMs: A stream of electrons is formed and

accelerated toward the specimen using a positive electrical potential

This stream is confined and focused using metal apertures and magnetic lenses into

a thin, focused, monochromatic beam.

This beam is focused onto the sample using a magnetic lens

Interactions occur inside the irradiated sample, affecting the electron beam

These interactions and effects are detected and transformed into an image

Two different types of EMs:
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-Transmission Electron Microscope (TEM) wasfirst type of electron microscope to be developed and

is patterned exactly on the Light Microscope exceptthat a focused beam of electrons is used instead of

light to see through the specimen

-It was developed by Max Knoll and Ernst Ruska inGermany in 1931

-The first Scanning Electron Microscope (SEM)debuted in 1942 with the first commercial instruments

around 1965


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TRANSMISSION ELECTRON MICROSCOPE(TEM)

-What you can see with a light microscope is limitedby the wavelength of light. TEMs use electrons as

"light source" and their much lower wavelength

makes it possible to get a resolution a thousand

times better than with a light microscope.
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HOW IT WORKS

-A "light source" at the top of the microscope emits theelectrons that travel through vacuum in the column of

the microscope.

-Instead of glass lenses focusing the light in the lightmicroscope, the TEM uses electromagnetic lenses to

focus the electrons into a very thin beam.

-The electron beam then travels through the specimen

you want to study.


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-Depending on the density of the material present,some of the electrons are scattered and disappear

from the beam.

-At the bottom of the microscope the unscattered

electrons hit a fluorescent screen, which gives rise to

a "shadow image" of the specimen with its differentparts displayed in varied darkness according to their

density.

-The image can be studied directly by the operator orphotographed with a camera.


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IMAGES FROM (TEM)

Leaf from green plants


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Sperma head of stick insect


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SCANNING ELECTRON MICROSCOPE (SEM)

The scanning electron microscope (SEM) is a type

of electron microscope that images the samplesurface by scanning it with a high-energy beam of

electrons in a raster scan pattern.
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HOW IT WORKS


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-The SEM is an instrument that produces a largely

magnified image by using electrons instead of light to

form an image.

-A beam of electrons is produced at the top of the

microscope by an electron gun.

-The electron beam follows a vertical path through

the microscope, which is held within a vacuum. The

beam travels through electromagnetic fields and

lenses, which focus the beam down toward thesample.


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-Once the beam hits the sample, electrons and X-

rays are ejected from the sample.

-Detectors collect these X-rays, backscattered

electrons, and secondary electrons and convert them

into a signal that is sent to a screen similar to a

television screen. This produces the final image.


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SAMPLE PREPARATION


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Samples have to be prepared carefully to withstand thevacuum inside the microscope.

Biological specimens are dried in a special manner thatprevents them from shriveling. Because the SEMilluminates them with electrons, they also have to be made

to conduct electricity.


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Our SEM samples are coated with a very thin layer ofgold by a machine called a sputter coater.


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IMAGES FROM SEM

This spiny-headed worm uses the spines on its head

to attach to the small intestine of a fish.
http://www.mos.org/sln/SEM/worm.gif


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Mosquito head
http://www.mos.org/sln/SEM/mhead.gif


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THE END

Microscope Chap 5

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