YOU NEED TO KNOW THIS STUFF Total Magnification In order to find total magnification you need to multiple the ocular lens by the objective Total Mag Ocular Lens x Objective Ex Ocular 10x Objective 40x ID: 549527
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Slide1
Measuring with a Microscope
YOU NEED TO KNOW THIS STUFF!Slide2
Total Magnification
In order to find total magnification you need to multiple the ocular lens by the objective.
Total Mag = Ocular Lens x ObjectiveEx.) Ocular = 10x, Objective = 40x Total Mag = 400xSlide3Slide4
Practice
Ocular = 15x, Objective = 30x, Total Mag =?
Ocular = ?, Objective = 40x, Total Mag = 400xOcular = 10x, Objective = ?, Total Mag =200xAnswers:450x10x20x Slide5
Field of View
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.Slide6
Field of View
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. Slide7
Field of View - Conversion
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. Ex.) 1.3 mm becomes 1300 micrometers
Once you get the field of view, you can then estimate the size of specimens. If the FOV was 1300 micrometers and something took up ½ the FOV, we could estimate it’s size to be 650 micrometersSlide8Slide9
Converting millimeters to micrometers
Multiply by 1000
Move the decimal point 3 places to the rightVideoSlide10
Calculating Specimen Size
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.
We can use the width of the field of view under low power and the relationship between the low & high power magnifications to mathematically calculate the width of the field of view under high powerSlide11
Calculating Specimen Size
First of all memorize this :
When switching from low to high power, the area in the field of view gets smaller & darker. 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 powerSlide12
When switching from low to high power…
You will see
LESS of your sample…it’s like zooming in on a pictureYou will see MORE detailThe diameter of the field of view DECREASES
The field of view gets DARKERSlide13Slide14Slide15
Example #1
ocular power = 10x
low power objective = 20x high power objective = 50x a) What is the highest magnification you could get using this microscope ? b) If the diameter of the low power field is 2 mm, what is the diameter of the high power field of view in mm? in micrometers ?
c) If 10 cells can fit end to end in the low power field of view, how many of those cells would you see under high power ? Slide16
Answer to A
ocular power = 10x
low power objective = 20x high power objective = 50x a) What is the highest magnification you could get using this microscope ? 500x Ocular x
high power = 10 x 50 = 500. (We can only use 2 lenses at a time, not all three.) Slide17
Answer to B
b) If the diameter of the low power field is 2 mm, what is the diameter of the high power field of view in mm ?
.8 mmThe ratio of low to high power is 20/50. So at high power you will see 2/5 of the low power field of view (2 mm). 2/5 x 2 = 4/5 = .8 mm in micrometers ? 800 micrometers
To convert mm to micrometers, move the decimal 3 places to the right (multiply by 1000). .8 mm x 1000 = 800 micrometers Slide18
Answer to C
c) If 10 cells can fit end to end in the low power field of view, how many of those cells would you see under high power ?
4 cells. We can answer this question the same way we go about "b" above. At high power we would see 2/5 of the low field. 2/5 x 10 cells = 4 cells would be seen under high power. Slide19
Example #2
ocular power = 10x
low power objective = 10x high power objective = 40x The diagram shows the edge of a millimeter ruler viewed under the microscope with the lenses listed above. The field shown is the low power field of view. a) What is the approximate width of the field of view in micrometers ?
b) What would be the width of the field of view under high power ? c) If 5 cells fit across the high power field of view, what is the approximate size of each cell ? Slide20
Answer to A
a) What is the approximate width of the field of view in micrometers ?
3500 - 3800 micrometers Each white space is 1 mm. We can see approximately 3 1/2 (or so) white spaces. That is equivalent to 3.5 mm, which converts to 3500 micrometers. Any answer in the range above would be OK. Slide21
Answer to B
b) What would be the width of the field of view under high power ?
875 micrometers The ratio of low to high power for this microscope is 10/40 or 1/4. So, under high power we will see 1/4 of the low power field of view. 1/4 x 3500 micrometers (from "a" above) = 875 micrometers. Slide22
Answer to C
c
) If 5 cells fit across the high power field of view, what is the approximate size of each cell ? 175 micrometers If 5 cells fit in the high power field of view (which we determined is 875 micrometers in "b"), then the size of 1 cell = 875/5 = 175 micrometers. Slide23
Review Questions!
1.) 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.) 2.) What about focusing ? How do you do that ?
Start on low power and turn the COARSE FOCUS knob until the slide looks clearOnce it is focused under low power you can move to the next objective and refocus if necessaryUnder high power only use the FINE FOCUS knob to focus in on your specimen Slide24Slide25Slide26
What Do The Parts Do?
1.the lens you look through, magnifies the specimen
ocular (eyepiece)2. supports the microscope
base3. 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
ArmSlide27
What Do The Parts Do?
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
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