Microscopy for Microbiology: A Primer

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An Overview of Microscopy
By BugLady
Unless otherwise stated, all micrographs and diagrams
are the author’s own work.
Jul-...

Microscopy
Resolution and magnification
Different types of light microscopy:
principle and use
Electron microscope: princi...

Principles of Light Microscopy
Light passes through specimen and through a
series of magnifying lenses
Important factors i...

Principles of Light Microscopy:
Magnification
Compound Microscope : microscope has two
magnifying lenses
 Lenses include ...

Principles of Light Microscopy: Resolution
Resolving power is defined as
the minimum distance existing
between two points ...

Principles of Light Microscopy: Resolution
Resolution
o Resolution depends on the quality of lenses
and wavelength of illu...

Principles of Light Microscopy Contrast
Reflects the number of visible shades in a
specimen
Higher contrast achieved for m...

Microscopy: The Instruments
Two lenses in the compound microscope
o Ocular lens (x10)
o Objective lens (x4, x10, x40, x100...

Label the Microscope
Arm
Base
Clamp
Condenser
Coarse
adjustment
Diaphragm
Fine adjustment
Light control
Jul-14 9
Light sou...

$Microscopy: The Instruments Refractive index is the light-bending ability of a medium. The light may bend in air so much t...$

Principles of Light Microscopy
Dark-Field Microscope
o Reverse image
 Specimen appears bright
on a dark background
Like ...

Electron Microscopy
Resolution is a function of wavelength:
the shorter the wavelength the higher
the resolution.
Uses ele...

Principles of Electron Microscopy
Uses electromagnetic lenses, electrons and
fluorescent screen to produce image
Resolutio...

Disadvantages of Electron Microscope
No true color
Artifacts
Large depth of field
Destroys sample
Jul-14 14

Staining
Live or unstained cells have little contrast with
the surrounding medium.
Cells are stained with dyes to make the...

Preparation of Specimens for
Light Microscopy
A smear is a thin film of a microbial liquid
suspension on a slide.
A smear ...

Preparing Smears for Staining
Stains consist of a positive ion and
negative ion.
In a basic dye, the chromophore is a
cati...

Simple Stains
Bacteria cell surfaces are slightly negatively charged
and basic dyes are used as stains.
Use of a single ba...

Differential Stains: Gram Stain
The Gram stain classifies bacteria into Gram-positive and
Gram-negative bacteria.
The Gram...

Differential Stains: Gram Stain
Jul-14 20
Color of
Gram + cells
Color of
Gram – cells
Primary stain: Crystal violet Purple...

Gram Stain Procedure Animation
This animation shows all the steps involved
in the Gram stain.
http://www.medschool.lsuhsc....

Differential Stain: Acid-fast Stain
Used to stain organisms that
resist conventional staining
Used to stain Mycobacterium
...

Special Stains
Stain specific structures in the bacterial
cell
Endospore stain
o Staining enhances endospore
o Uses heat t...

Morphology of Prokaryotic Cells
Prokaryotes exhibit a
variety of shapes
o Most common
 Coccus
Spherical
 Bacillus
Rod ...

Morphology of Prokaryotic Cells
Prokaryotes exhibit a variety
of shapes
o Coccobacillus: Short round rod
o Vibrio: Curved ...

Morphology of Prokaryotic Cells
Division along a single plane may result in pairs or
chains of cells
Pairs = diplococci
o ...

Morphology of Prokaryotic Cells
Division along two or three
perpendicular planes form
cubical packets
Example: Sarcina gen...

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