LABORATORY LESSON 1: MICROSCOPY
CHARIELLE MAE P. VINLUAN, RMT,WMA
OBJECTIVES:
1. To introduce students of one of the chief scientific tools in microbiology
2. To impress students with the importance of proper care and maintenance of such a precision instrument
COMPOUND MICROSCOPES
• Constructed by Zaccharias and Hans Janssen
• Consisted of an object lens and an ocular lens which are separated by a body tube
• Produces a two-stage magnification
TYPES OF MICROSCOPES
A. LIGHT MICROSCOPE
• The light microscope is an instrument for visualizing fine detail of an object. It does this by creating a magnified image through
the use of a series of glass lenses, which first focus a beam of light onto or through an object, and convex objective lenses to
enlarge the image formed
• Uses visible light to detect small objects
Types of Light Microscope
a. Brightfield Microscope
• Most commonly used light microscope in a clinical laboratory
• Forms a dark image against a brighter background
• Kohler illumination
Parts of a Brightfield microscope
Eyepiece
Diopter adjustment ring
Arm
Revolving nosepiece
Objective lens
Mechanical stage
Condenser
Iris diaphragm
Illuminator
Base
Mechanical stage adjustment
knobs
Coarse adjustment knob
Fine adjustment knob
TERMINOLOGIES
a. MAGNIFICATION
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• Process of enlarging an object’s size without affecting its actual size
• Total magnification= Ocular x objective magnification
b. FOCAL POINT
• Specific point at which parallel rays of light converge after passing through a lens
c. FOCAL LENGTH
• The distance from the lens center to a point where parallel rays are focused on the optical axis
d. RESOLUTION OR RESOLVING POWER
• Ability of a lens to distinguish two points or objects as separate
e. WORKING DISTANCE
• Distance between the objective and the coverslip on the slide
f. NUMERICAL APERTURE
• Number that indicates the resolving power of a lens system
b. Darkfield microscope
• The condenser does not allow light to pass directly through the specimen but directs the light to hit the specimen at an oblique
angle
• Organism will appear extremely bright against a darkfield
• Used to view spirochetes (Treponema pallidum)
c. Fluorescent Microscope
• Allows visualization of naturally fluorescent microorganisms or those stained by a fluorescent dye (antibody) or fluorophores
• Involves the excitation of fluorochromes using lights
o Excitation filter
o Barrier filter
• Dyes: Acridine orange, auramine, rhodamine, calcofluor white, FITC
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d. Phase-contrast microscope
• Light beams are deflected by different thickness of the object. The light beams are reflected a second time when they strike
the objective.
• It permits a detailed examination of internal structures in living organism
• Can be used to examine unstained living cells
• Brightfield microscope can be converted into a phase-contrast
• Areas of the specimen appear light and dark with haloes of various intensities
B. ELECTRON MICROSCOPE
• Is one of the most advanced and important types of microscopes highest magnifying capacity
• Electrons are used to illuminate the tiniest particles
• More powerful tool than light microscopes
Types of Electron Microscope
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