Lab 2

Microscope
 Objectives

■Part A – Microscope Overview

■Part B – Magnification vs. Resolving Power
■Part C – Using the Microscope
■Part D – Practice – letter “e”
■Part E – Measurements

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 Types of Microscopes

• Compound Light Microscope (transmitted light)

• Dissecting or Binocular Light Microscope (reflected light)
• Transmission Electron Microscope (TEM) (transmitted
electron beam)
• Scanning Electron Microscope (SEM) (reflected electron
beam)

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 The Light Microscope/Compound

■ Several types
– bright-field microscope
– dark-field microscope
– phase-contrast microscope
– fluorescence microscope
■ Are compound microscopes
– image formed by action of  2 lenses

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 Why Use A Microscope?

■ Magnification – enlarge the objects

■ Resolution – make objects clearer

■ Important tool of biologists

■ Used to study cells, tissues, and microorganisms

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 Brightfield Microscope

 allows light rays to pass directly to the eye without being

deflected by an intervening opaque plate in the condenser

 produces a dark image against a brighter background

 has several objective lenses

 typical microscope used by college students

 parfocal microscopes remain in focus when objectives are

changed

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 Total Magnification

■ A simple calculation
■ Simply multiply the ocular magnification by the
magnification of the objective you are using
– Ocular magnification (15x) X Objective magnification (45x) = 675x

■ magnification of each ocular and objective lens is usually

written on it

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 Compound Microscope

 Total magnification
 Product of the magnifications of the ocular
lenses and the objective lenses

45x (objective) X 10x (eye piece)=

450x

 Resolution
 ability of a lens to distinguish one point from
another at very small distances(max ~ 0.2 µm)
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Objective Lens

■ Example: 10x / 0.25

■ 10x: means that it magnifies the

object 10 times.

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 Improving Resolving Power
 Blue filter - should be placed over the light source because the shorter 
of the resulting light will provide maximum resolution

 Condenser - should be kept at the highest position that allows the

maximum amount of light to enter the objective lens and therefore limit
the amount of light lost due to refraction

 Diaphragm- should not be stopped down too much. While closing the
diaphragm improves the contrast, it also reduces the numerical aperture

 Immersion oil - used between the slide and the 100 objective lens
 special oil that has the same refractive index as glass
 when placed between the specimen and objective lens, the oil forms
a continuous lens system that limits the loss of light due to
refraction

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 Magnification

 Compound Light Microscope: around 1500x

 Transmission Electron Microscope: around

250,000x (biologists usually use less)

 Although you could easily make lenses for light

microscopes to magnify more, the images would be
blurry due to the lower resolution

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Care of The Instrument
TRANSPORT

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Carrying a Microscope

A B
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Carrying a Microscope

A B
• Carry it with both hands
• Keep it vertically and not tilted as the oculars can
fall off
• Clean the optical system before and after usage:
• Ocular lens
• Objectives
• Condenser lens

• Use only optical lens tissue

• Never remove parts of the microscope
• At the end of work place 4X or 10X objective in
place (which ever is the smallest)
• Cover microscope with cover

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 Lens Care
 Keeping the lenses clean of dust, oil, clean of
contaminants is very important

 Cleaning Tissues – oil-lint free special for lenses

 Solvents – green soap with warm water, xylene,

alcohol and acetone

 Oculars – check weather is clean -rotate it between

the thumb and forefinger as you look through the
microscope

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 Lens Care

• Use lens paper on all glass parts of the microscope.

• Clean oil immersion lens with chemicals provided by
your instructor
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1. Clutter – maintain a clean work desk

2. Electric Cord

3. Lens care (clean before and after lab); remove

any trace of immersion oil

4. Dust protection – use dustcovers

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 Microscope Components
 Head (supports the 2 sets of magnifying lenses)
 Ocular – is the eyepiece (common 10X magnification)
 Interpupillary distance adjustable
 Field of view (has pointer)
 Framework – includes arm and base
 Stage with stage adjustment with knobs
 Light source
 Focusing Knobs - fine and coarse used to bring objects into
focus
 Condenser lens - focus the light from the lamp and it can
be adjusted up and down
 Iris diaphragm – controls the width of the circle of light
(amount of light passing )
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 4X or 10X – low power objective or scanning
objective

 40X – high power objective; it is used dry

 100X – immersion oil

 Objective are PARFOCAL

 Microscope can have:

 10X, 43X, 97X

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Compound Microscope

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 Lens Systems in Compound Light
Microscope
1. Oculars (eyepieces): provide some magnification, focus
image at eye (can be 1 or 2 of them); has magnification of 10X
- Modern microscope – binocular

2. Objectives located on rotating nosepiece, provide

magnification and resolving power (may be 3 to 6 of them)

- 10X (low-power), 40X (high-dry), 100X (oil immersion);

some have even 4X (for rapid scanning)
3. Condenser located under stage, focuses light on the
specimen
- It may also contain an iris diaphragm that controls the size
of the cone of light entering the condenser

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 Lens Systems in Compound Light
Microscope
1. Oculars (eyepieces): provide some magnification, focus
image at eye (can be 1 or 2 of them); has magnification of 10X
- Modern microscope – binocular

2. Objectives located on rotating nosepiece, provide

magnification and resolving power (may be 3 to 6 of them)

- 10X (low-power), 40X (high-dry), 100X (oil immersion);

some have even 4X (for rapid scanning)
3. Condenser located under stage, focuses light on the
specimen
- It may also contain an iris diaphragm that controls the size
of the cone of light entering the condenser

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 Parts of the Microscope
• Ocular • Fine adjustment knob
• Body tube • Stage
• Nosepiece • Stage clips
• Objective lenses • Aperture
(10, 40, 100x) • Diaphragm
• Arm
• Light
• Course adjustment
• Base
knob
 Parfocal Microscope

 image will remain in focus when changing from a lower-

power objective lens to a higher-power lens

 minimal focusing should be necessary with the fine focus

adjustment

http://www.youtube.com/watch?v=f7KlFSgdUGU

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Putting It Away

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