MICROSCOPY

INTRODUCTION
Microscopy is defined as the use of a microscope
to magnify objects too small to be visualized with
the naked eye so that their characteristics are
readily observable

APPLICATION OF MICROSCOPY
IN DIAGNOSTIC MICROBIOLOGY
Rapid preliminary organism identification
Rapid final identification of certain organisms
Detection of different organisms +nt in same
specimen
Detection of organism not cultivated in lab.
Provide preculture information

TERMINOLOGY
Magnification:

The degree of enlargement

It is expressed as the number of times the length,
breadth or diameter, but not area, of the object is
multiplied
Magnification is effected in two stages objective
lenses & eye pieces lenses
The magnifying power of an objective

 3 objective - a. low power dry X 10

b. high power dry X 40
c. oil immersion X 100
Eye piece magnification X 10
Over all magnification of the
microscope X 100, X 400 & X 1000

Contd..
Resolution :
The minimum distance between two visible
bodies at which they are seen as separate & not
in contact with one another

TYPES OF MICROSCOPE
BRIGHT FIELD( LIGHT) MICROSCOPE:
PRINCIPLE:
Visible light passed through specimen &
through series
of lenses that reflect light in a manner that
results in magnification of organisms +nt in
specimen

Darkground microscope
Principle
The specimen is illuminated only by
rays of light so oblique that unless
they are scattered by objects

Darkground microscope
PRINCIPLE:

Phase contrast
microscope
Objects are made to appear in darkgrey contrast against a bright
background by causing direct &
diffracted rays from them, which
differ in their wave phase, to
recombine in such a way that they
interfere with each other & so reduce
the light intensity in the area of the
image corresponding to the object

Phase contrast microscope

PRINCIPLE:

Fluorescence microscope
When certain materials, e.g. oil or fat
droplets, or certain dyes are exposed
to ultra violet radiation, they convert
this invisible, short wave radiation
into the longer wavelength radiation
of visible light, & so become
luminous & are said to fluorescence

Fluorescence microscope
PRINCIPLE:
- Fluorescent dye is used to stain the smear

Electron Microscope
Two types:
- SEM (Scanning Electron Microscope)
- TEM (Transmission Electron Microscope)

SEM (Scanning Electron

Microscope)

(TEM)Transmission Electron
Microscope

MICROSCOPY IN DIAGNOSTIC
MICROBIOLOGY
Organism
Bright field
Fluorescence
Dark field
Electron
Group
microscope microscope
microscope
microscope
Bacteria

Fungi +

Parasites

Viruses

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