CHAPTER 5 MICROSCOPE

-Device that can be used to observe the objects that are too

small to be seen by the naked instrument or unaided eye.

-The science of investigating small objects using such an

instrument 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 microscope allowed people to see things no human had ever seen before.

-The first useful microscope was Netherlands between 1590 and 1608

developed

in

the

-There are THREE types of microscopes:

- Compound Light Microscope - Electron Microscope

Transmission Electron Microscope (TEM) Scanning Electron Microscope (SEM)

COMPOUND LIGHT MICROSCOPE

-The term light refers to the method by which light transmits the image to your eye. Compound deals with the microscope 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.

COMPOUND LIGHT MICROSCOPE

Diagram Showing Light Traveling Through The Microscope

Magnifying Objects/ Focusing Image:

-When viewing a slide through the microscope make sure that the stage is all the way down and the 4X scanning objective is locked into place. -Place the slide that you want to view over the aperture and gently move the stage clips over top of the slide to hold it into place. -Beginning with the 4X objective, looking through the eyepiece making sure to keep both eyes open (if you have trouble cover one eye with your hand) slowly move the stage upward using the coarse adjustment knob until the image becomes clear.

-To magnify the image to the next level rotate the nosepiece to the 10X objective. While looking through the eyepiece focus the image into view using only 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 a slight turn of the fine adjustment knob to complete this task.

Total Magnification:
-To figure the total magnification of an image that you are 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.

IMAGES FROM COMPOUND LIGHT MICROSCOPE

Elodea 40X

Elodea 100X

Elodea 400X

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 very important 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 for your hair

Care and Handling Transporting:

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

Handling & Cleaning: Never touch the lenses with your fingers. Your body produces an oil that smudges the glass. This oil can even etch the glass if left on too long. Use only LENS PAPER to clean the glass. TOILET PAPER, KLEENEX, AND PAPER TOWELS HAVE FIBERS THAT CAN SCRATCH THE LENSES.
Storage:
When you are finished with your "scope" assignment, rotate the nosepiece so that it's on the low power objective, roll the nosepiece so that it's all the way down to the stage, then replace the dust cover. DON'T FORGET TO USE PROPER TRANSPORTING TECHNIQUES!

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 white circle of light.

6. Place a slide on the stage. Center the specimen over the opening on the stage. Use the stage clips to hold the slide in place.

6. Place a slide 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 highpower objective lens clicks into place. Make sure the lens does not hit the slide. 10. Looking through the eyepiece, turn the fine adjustment knob until the specimen comes into focus.

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 of the interior structures of organic cells (nucleus, mitochondria..) -This required 10,000 plus magnification which was just not possible using Light Microscope What is Electron Microscopes? Electron Microscopes are scientific instruments that use a beam of highly energetic electrons to examine objects on a very fine scale.

How do Electron Microscopes Work? 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: Transmission Electron Microscope Scanning Electron Microscope

-Transmission Electron Microscope (TEM) was first type of electron microscope to be developed and is patterned exactly on the Light Microscope except that a focused beam of electrons is used instead of light to see through the specimen
-It was developed by Max Knoll and Ernst Ruska in Germany in 1931 -The first Scanning Electron Microscope (SEM) debuted in 1942 with the first commercial instruments around 1965

TRANSMISSION ELECTRON MICROSCOPE(TEM)

-What you can see with a light microscope is limited by 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.

HOW IT WORKS
-A "light source" at the top of the microscope emits the electrons that travel through vacuum in the column of the microscope. -Instead of glass lenses focusing the light in the light microscope, 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.

-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 different parts displayed in varied darkness according to their density. -The image can be studied directly by the operator or photographed with a camera.

IMAGES FROM (TEM)

Leaf from green plants

Sperma head of stick insect

SCANNING ELECTRON MICROSCOPE (SEM)

The scanning electron microscope (SEM) is a type of electron microscope that images the sample surface by scanning it with a high-energy beam of electrons in a raster scan pattern.

HOW IT WORKS

-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 the sample.

-Once the beam hits the sample, electrons and Xrays 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.

SAMPLE PREPARATION

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

Biological specimens are dried in a special manner that prevents them from shriveling. Because the SEM illuminates them with electrons, they also have to be made to conduct electricity.

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

IMAGES FROM SEM

This spiny-headed worm uses the spines on its head to attach to the small intestine of a fish.

Mosquito head

THE END