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DEFINITION
- A microscope (Greek: micron = small and scopos = to look)
- MICROSCOPE: Is an instrument for viewing objects that
are too small to be seen by the naked eye
- Microscope must accomplish three tasks:
* Magnification
* Resolution
* Contrast
3
MAGNIFICATION
Degree of enlargement
No of times the length, breadth or diameter, of an
object is multiplied.
RESOLUTION
Ability to reveal closely adjacent structural
details as separate and distinct
LIMIT OF RESOLUTION (LR): The min distance between two visible bodies at
which they can be seen as separate and not in contact with each other
Types of microscope
Resolving power
Compound Microscope
200 nanometers
10 nanometers
0.2 nanometers
5
Contrast
- Giving the details visible to the eye, camera, or other imaging
device
HISTORICAL
BACKGROUND
nd
st
Century: Romans experiment with glass and observe objects larger when
viewed through glass
1590: Hans Janssen and his son Zacharias Janssen, mount two lenses in a
tube to create first compound microscope
1609: Galileo Galilee develops compound microscope with concave and convex
lens
attention of biologists.
10
Safety
IS FIRST
The following symbols are found on the microscope. Study the meaning of the symbols
and always use the equipment in the safest possible manner
TYPES OF MICROSCOPE
- Simple microscope
- Compound microscope
- Electron Microscope
- Phase Contrast
Microscope
- Dark Ground Microscope
- Fluorescent Microscope
- Others
SIMPLE MICROSCOPE
More than five hundred years ago, simple glass magnifiers were
developed. These were convex lenses (thicker in the center than the
periphery). The specimen or object could be focused by use of the magnifier
placed between the object and the eye. These simple microscopes, along
with the cornea and eye lens, could spread the image on the retina by
magnification through increasing the visual angle on the retina.
Convex Lenses are curved glass used to make microscopes (and glasses etc.)
Convex Lenses bend light and focus it in
one spot
The image produced by such a magnifier, held close to the observers eye,
appears as if it were on the same side of the lens as the object itself. Such an
image, seen as if it were ten inches from the eye, is known as a virtual image.
These magnifiers had severe limitations in specimen positioning, illumination,
lens aberrations, and construction.
Simple magnifier. A simple magnifier uses a single lens system to enlarge the object in
one step
COMPOUND
MICROSCOPE
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Parts of Microscope
i. Optical component
ii.Mechanical component
iii.Electrical component
Optical component
OBJECTIVE LENS
1. Its basic function is to gather the light passing through the specimen
and then to project an accurate, real, inverted IMAGE of the specimen
up into the body of the microscope.
2. Other related functions of the objective are simply to magnified the
specimen with a given magnification rate i.e 4x,10x ,40x and 100x
magnification power
Scan - 4X
Low Power - 10X
TYPES
FBEG - OIL
ABCD - AIR
OIL
C GLASS
B
F
25
1.
2.
3.
4.
5.
Working distance
Objective
Magnification
Working
Distance
4X
18.5
10X
10.6
40X
0.6
100X
0.13
EYE PIECE
1. Its basic function is to look at the
focused, magnified real image
projected by the objective.
2. They are usually 10x,15x,40x and
45x
- TYPES
(a) Monocular(Huygenian)
(b) Binocular (Ramsden)
(c) Trinocular (Compensating)
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10x
40x
SUBSTAGE CONDENSER
1. Its basic function is to gather the light coming from the light source and to
concentrate(focus) that light in a collection of parallel beams onto the
specimen.
COLLECTOR LENS.
This lens is placed in front of the light source. Its function is to project an
image of the light source onto the plane of the condensers aperture
diaphragm. In some instruments a diffusion or frosted filter is placed just
after the collector lens (side closer to the specimen) in order to provide
more even illumination.
TUBE LENS
Its function is to gather the parallel rays of light projected by the objective
and bring those rays to focus at the plane of the fixed diaphragm of the
eyepiece.
Body Tube
The body tube holds the objective lenses
and the ocular lens at the proper distance
Diagram
Nose Piece
The Nose Piece holds the objective lenses
and can be turned to increase the
magnification
Diagram
Stage Clips
These 2 clips hold the slide/specimen in
place on the stage.
Diagram
CONDENSER HOLDER.
This holder receives any one of several types of condenser
Diaphragm
The Diaphragm controls the amount of light
on the slide/specimen
Diagram
Arm
Used to support the microscope when
carried. Holds the body tube, nose piece
and objective lenses
Diagram
Stage
Supports the slide/specimen
Diagram
Diagram
Diagram
Base
Supports the microscope
Diagram
48
* switch
* Rheostat
* lamp (halogen or incandescent)
* fuse
* Electronic sub system
Light Source
Projects light upwards through the
diaphragm, the specimen and the lenses
Some have lights, others have mirrors
where you must move the mirror to reflect
light
Diagram
Operational Principle
Light
Knowledge of the behavior of light and the effects resulting when light passes from
air through a glass convex lens and out into air again is fundamental to the
understanding of image formation. When light passes from air into a convex lens,
the speed of light is slowed. The various colors, differing in wave length, are slowed
at different rates (dispersion). This bending (refraction) effect differs for different
colors. Those rays which strike the central area of the lens at a perpendicular
emerge unrefracted. Light passing through the other parts of the convex lens are
refracted or bent. The blue rays are bent more than thegreen rays, more than
the red rays.
When white light passes through convex lenses of objectives, eyepieces, or
condensers, two main kinds of aberrations may occur; chromatic aberration and/or
spherical aberration. These aberrations can be corrected in the design of the
lenses.0
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General Maintenance
Preventive Maintenance
Microscope frame
Observation Tube optical axis
Observation Tube left/right axis
Observation Tube revolving axis
Observation Tube parfocality
As needed
General Maintenance
General Maintenance
General Maintenance
Cleaning the eyepieces
*When cleaning oil immersion objective, first wipe objective with a dry lens tissue
General Maintenance
General Maintenance
General Maintenance
Preparing for Inspection
General Maintenance
Inspection Sheet
Use the CX21 inspection sheet
and check the electrical unit,
mechanical and optical
performance
General Maintenance
Inspection: Checking for Dust
Image influence caused by dirt on each component
General Maintenance
Inspection: Checking for Dust
Note: If dirty portion
can not be
identified, it is
assume that the
internal lens and
prism are
contaminated.
General Maintenance
Inspection: Checking Lenses
General Maintenance
Inspection: Checking Lenses
General Maintenance:
Checking Left/Right Alignment
Place the cross eyepiece into the right
sleeve
Place a slide with an image with a
recognizable reference point (e.g. center
of a spiral), secure it on stage
Align the reference point in the center of
the cross of the eyepiece (looking
through right eyepiece) by moving the
stage with the control knob
General Maintenance:
Checking Left/Right Alignment
Cont.
General Maintenance:
Checking Left/Right Alignment
Cont.
General Maintenance:
Checking Left/Right Alignment
Common Failures
Electrical/Power Failure
Dirty components
Eyepiece
Objectives
Filters
Not in focus
Brightness not adjusted properly
Objective not rotated fully into secured position
Troubleshooting
What is troubleshooting?
A logical, systematic process that is able to determine "what" happened, "why" it
happened and a method to develop effective fixes for the "why" it happened.
Why do we Troubleshoot?
To prevent the equipment and machinery from repeat incidents and failures
5 Steps of Troubleshooting
o Preparation
o Step 1 Observation
o Step 2 Define Problem Area
o Step 3 Identify Possible Causes
o Step 4 Determine Most Probable Cause
o Step 5 Test and Repair
o Follow-up
Preparation
o Before you begin to troubleshoot any piece of
equipment, you must be familiar with safety rules and
procedures for working on electrical equipment.
Be Safe!
Turn off power, ground yourself, and use gloves when
appropriate
Step 1: Observation
o What Happened?
o Most faults provide obvious clues as to their cause.
o Through careful observation and a little bit of reasoning,
most faults can be identified as to the actual component
with very little testing.
were
identified either by past history or by the person that
reported the problem.
only the
part(s). If faulty, it could cause the symptoms that the
equipment is experiencing.
o Starting with the whole circuit as the problem area, take each
noted observation and ask yourself, "what does this tell me about
the circuit operation?".
o The following list shows the order in which you should check
components based on the probability of them being defective:
1. Components that burn out or have a tendency to wear out: i.e.
mechanical switches, fuses , relay contacts, and light bulbs.
(Remember, fuses burn out for a reason. You should find out why
before replacing them.)
2. Coils, motors, transformers, and other devices with windings:
These usually generate heat and with time, can malfunction.
3. Connections: especially screw or bolted type. Over time these
can loosen and cause a high resistance. In some cases this
resistance will cause overheating and eventually will burn open.
Connections on equipment that is subject to vibration, are
especially prone to coming loose.
4. Defective wiring: Pay particular attention to areas where the
wire insulation could be damaged causing short circuits. Do not
rule out incorrect wiring, especially on a new piece of equipment.
Follow-Up
Not an official step of the troubleshooting process, but it
should be done once the equipment has been repaired and
put back in service.
Troubleshooting
Troubleshooting
Electrical Unit
Troubleshooting
Electrical Unit
After verifying the light bulb is not the issue,
check that the microscope is plugged in and
that outlet has power(use another device
you know to be working to validate this)
Verify the chord shows no obvious damage
Verify the fuse is intact
Troubleshooting
Troubleshooting
Troubleshooting
Stage falls down spontaneously or an infocus image quickly moves out of focus
Troubleshooting: Backlash
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Dust
Protect the microscope from dust- make sure you use a dust
cover.
Source: www.funsci.com/fun3_en/dust/dust.htm
2.
Humidity
Using a plastic dust jacket (like the one supplied with the
scope) is a bad idea.
SOLUTION: Buy some thick calico and use the plastic cover as a
template to make dust covers that are washable and can
breathe. This prevents condensation inside the cover and the
development of ideal conditions for fungal growth (warm and
wet)
3.
Water
Summary