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The Microscope
• That gizmo pictured to the
left is a BIG deal. It literally
opened up worlds of organisms
and information to scientists.
It's importance in the history
of medicine and our
understanding of disease
should not be underestimated.
That gizmo is a compound
light microscope.
For you, the biology student,
it is perhaps the most
important tool for you to
understand. You should be
able to :
1. name all of its parts and
describe the function of each
2. explain how to carry the
thing, properly prepare a
slide, & focus correctly
3. calculate total
magnification
4. estimate the size of a
specimen being observed
What the parts do!
1.the lens you look through, magnifies the specimen ocular (eyepiece)
2. supports the microscope base
3. holds objective lenses nosepiece
4. magnify the specimen (2) high power objective lens
low power objective lens
5. supports upper parts of the microscope, used to carry the microscope
Arm
6. used to focus when using the high power objective fine focus knob
7. where the slide is placed stage
8. regulates the amount of light reaching the objective lens diaphragm
9. used to focus when using the low power objective coarse focus knob
10. provides light light source
11. hold slide in place on the stage stage clips
Important 'Scope Vocab :
magnification \mag-ne-fe-'ka-shen\ n 1. apparent enlargement of an object 2.
the ratio of image size to actual size
A magnification of "100x" means that the image is 100 times bigger than the
actual object.
resolution \rez-e-loo-shen\ n 1. clarity, sharpness 2. the ability of a microscope
to show two very close points separately
OK, well. There are a few other tidbits about the compound microscope you
should remember :
1. Why is called a "compound" light microscope ?
"Compound" just refers to the fact that there a two lenses magnifying the
specimen at the same time, the ocular & one of the objective lenses.
2. If two lenses are always magnifying the specimen
(see #1), how do you figure out the total magnification being used ?
You multiply the power of the ocular and the power of the objective being used.
total mag. = ocular x objective
For example, if the ocular is 10x and the low power objective is 20x, then the
total magnification under low power is 10 x 20 = 200x.
Easy, ain't it ?
3. How do you carry one of those things ?
With two hands, one holding the arm & the other under
the base. Kinda like a football. (They're expensive, we
don't want to drop 'em.)
4. What about focussing ? How do you do that ?
Here's what I suggest. Once you have your slide in
place on the stage, make sure the low power objective
(the shortest objective lens) is in position & turn the
coarse focus until the lens is at a position closest to the
stage. Set the diaphragm to its largest opening (where it
allows the most light through). Then, while looking
through the ocular, begin to slowly turn the coarse
focus. Turn slowly & watch carefully. When the
specimen is focussed under low power, move the slide
so that what you want to see is dead-center in your field
of view, & then switch to a higher power objective. DO
NOT touch the coarse focus again --- you will break
something ! Once you are using a high power objective,
focus using the fine focus knob ONLY. Be sure to center
your specimen before switching to a higher power
objective or it may disappear.
MICROSCOPIC
MEASUREMENTS
Estimating Specimen Size
The area of the slide that you see when you look through a microscope is
called the "Field of View". If you know how wide your field of view is, you
can estimate the size of things you see in the field of view. Figuring out the
width of the field of view is easy --- all you need is a thin metric ruler.
By carefully placing a thin metric ruler on the stage (where a slide would
usually go) and focusing under low power, we can measure the field of view
in millimeters. Through the microscope it would look something like what
you see here on the left. The total width of the field of view in this example
is less than 1.5 mm. A fair estimate would be 1.3 or 1.4 mm.
(Relax, it's an estimate).
Now millimeters is a nice metric unit, but when we use a MICROscope we
tend to use MICROmeters. To convert from millimeters to micrometers,
move the decimal 3 places to the right. Our 1.3 mm estimate becomes
1300 micrometers.
Now we can get the ruler out of the way, prepare a slide, focus, and
estimate the size of things we see ! (Exciting, ain't it ?)
For example, if something we were looking at took up half of the field of
view, its size would be approximately 1/2 x 1300 micrometers = 650
micrometers. If something appeared to be 1/5 of the field of view, we would
estimate its size to be 1/5 x 1300 = 260 micrometers.
MICROSCOPIC
Calculating Specimen Size
MEASUREMENTS
Because the high power objective is so close to the stage, we can't
measure the width of the field of view under high power directly.
The ruler just doesn't fit between the objective & the stage. No
problem. We can use the width of the field of view under low power
(which we measure using the steps above) and the relationship
between the low & high power magnifications to mathematically
calculate the width of the field of view under high power.
First of all memorize this :
When switching from low to high power, the area in the field of view
gets smaller & darker. (You see a smaller area of the slide under
high power.) This is why centering what you want to see prior to
switching to high power is so important.
The fraction of the area seen under high power is the same as the
ratio of the low & high power magnifications.
For example : if the low power objective is 20x and the high power
objective is 40x, then under high power we will see 20/40 or 1/2 of
the area of the slide we saw under low power.
Example #1: