The Use and Care of Microscopes

THE USE AND CARE OF
MICROSCOPES
Presented by: Tom Anthony A. Tonguia, RMT

JMCFI – MLS Department
MLS 3A – Bacteriology Laboratory
OBJECTIVES
1. Identify and describe the different parts and functions of the bright-field
compound microscope;
2. Demonstrate how to properly use and care for the microscope; and
3. Describe the importance of the microscope in bacteriology laboratory.
OVERVIEW
• Microbiology, the branch of science that has so vastly extended and expanded
our knowledge of the living world, owes its existence to Anton van
Leeuwenhoek.
• Leeuwenhoek introduced the world to the existence of microbial forms of life.
• Microscopes are designated as either light microscopes or electron
microscopes.
ESSENTIAL FEATURES OF
MICROSCOPES
1. Brightfield Microscope
• This instrument contains two lens systems for magnifying specimens: the
ocular lens in the eyepiece and the objective lens located in the nose-piece.
• The specimen is illuminated by a beam of tungsten light focused on it by a
sub-stage lens called a condenser, and the result is that the specimen appears
dark against a bright background.
• Limitations:
• Absence of contrast between the specimen and the surrounding medium,
which makes it difficult to observe living cells
MICROSCOPES
2. Darkfield Microscope
• This is similar to the ordinary light microscope; however, the
condenser system is modified so that the specimen is not illuminated
directly.
• The con-denser directs the light obliquely so that the light is
deflected or scattered from the specimen, which then appears bright
against a dark background.
MICROSCOPES
3. Phase-Contrast Microscope
• Observation of microorganisms in an unstained state is possible with this
microscope.
• Its optics include special objectives and a condenser that make visible
cellular components that differ only slightly in their refractive indexes.
• As light is transmitted through a specimen with a refractive index different
from that of the surrounding medium, a portion of the light is refracted
(bent) due to slight variations in density and thickness of the cellular
components.
MICROSCOPES
4. Fluorescent Microscope
• This microscope is used most frequently to visualize specimens that are
chemically tagged with a fluorescent dye.
• The source of illumination is an ultraviolet (UV) light obtained from a high-
pressure mercury lamp or hydrogen quartz lamp.
• The ocular lens is fitted with a filter that permits the longer ultraviolet
wavelengths to pass, while the shorter wavelengths are blocked or
eliminated.
MICROSCOPES
5. Electron Microscope
• This instrument provides a revolutionary method of microscopy, with
magnifications up to one million.
• This permits visualization of submicroscopic cellular particles as well as
viral agents.
• In the electron microscope, the specimen is illuminated by a beam of
electrons rather than light, and the focusing is carried out by electromagnets
instead of a set of optics.
THEORETICAL PRINCIPLES OF
MICROSCOPY
1. Magnification
• Enlargement or magnification of a specimen is the function of a two-lens system; the
ocular lens is found in the eyepiece, and the objective lens is situated in a revolving
nose-piece.
• These lenses are separated by the body tube.
• The objective lens is nearer the specimen and magnifies it, producing the real image
that is projected up into the focal plane and then magnified by the ocular lens to
produce the final image.
MICROSCOPY
2. Resolving Power or Resolution

• Resolving power is the ability of a lens to show two adjacent objects as discrete
entities.
• When a lens cannot discriminate, that is, when the two objects appear as one, it has
lost resolution. Increased magnification will not rectify the loss, and will, in fact, blur
the object.
• The resolving power of a lens is dependent on the wave-length of light used and the
numerical aperture, which is a characteristic of each lens and imprinted on each
objective
MICROSCOPY
3. Illumination
• Effective illumination is required for efficient magnification and resolving power.
• The light is passed through the con-denser located beneath the stage.
• The condenser contains two lenses that are necessary to produce a maximum
numerical aperture.
• The height of the condenser can be adjusted with the con-denser knob.
• Excessive illumination may actually obscure the specimen because of lack of contrast.
USE AND CARE OF THE MICROSCOPE
• Once the microscope is placed on the laboratory bench,

observe the following rules:
1. Remove all unnecessary materials such as books, papers, purses,
and hats from the laboratory bench.
2. Uncoil the microscope's electric wire and plug it into an electrical
outlet.
3. Clean all lens system
• The following routine procedures must be followed to ensure correct and

efficient use of the microscope while focusing:
1. Place the microscope slide with the specimen within the stage clips on the fixed
stage. Move the slide to center the specimen over the opening in the stage directly
over the light source.
2. Rotate the scanning lens or the low power lens into position. While watching from
the side to ensure that the lens doesn't touch the specimen, turn the coarse focus knob
to move the stage as close as it can get to the lens without touching the lens.
3. Now, while looking through the ocular lens, turn the coarse focus knob carefully, and
slowly move the stage away from the lens until the specimen comes into vague focus
4. If this is the first specimen of the day, you should Kohler your microscope at this
point
• The following routine procedures must be followed to ensure correct and

efficient use of the microscope while focusing:
5. Routinely adjust the light source by means of the light source transformer setting,
and/or the iris diaphragm, for optimum illumination for each new slide and for each
change in magnification.
6. Our microscopes are parfocal, which means that when one lens is in focus, other
lenses will also have the same focal length and can be rotated into position without
further major adjustment
7. Once you have brought the specimen into sharp focus with a low-powered lens,
preparation may be made for visualizing the specimen under oil immersion.
8. During microscopic examination of microbial organisms, it is always necessary to
observe several areas of the preparation
MICROSCOPE CLEANING AND
MAINTENANCE
• To clean the exterior side of lenses, use a non-solvent cleaning solution

designed for cleaning optics or eyeglasses.
• First remove dust with a soft brush or can have compressed air.
• Then moisten a piece of dry lint-free tissue or lens paper and clean the lens
surfaces with a circular motion.
• Repeat with a second piece of paper moistened with lens solution if necessary.
• Repeat once again with dry lens paper until the lens is clean and dry.
• Use this same procedure for the eyepiece and objective lenses.

The Use and Care of Microscopes

Uploaded by

Copyright:

Available Formats

The Use and Care of Microscopes

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

The Use and Care of Microscopes

Uploaded by

Copyright:

Available Formats

THE USE AND CARE OF

Presented by: Tom Anthony A. Tonguia, RMT

2. Resolving Power or Resolution

• Once the microscope is placed on the laboratory bench,

• The following routine procedures must be followed to ensure correct and

• The following routine procedures must be followed to ensure correct and

• To clean the exterior side of lenses, use a non-solvent cleaning solution

You might also like