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Tools of the Laboratory

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Misty Wehling

on 16 July 2013

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Transcript of Tools of the Laboratory

Tools of the
Laboratory

Staining
Techniques

The 6 "I"s
Microscopes
All about the tools we use to
work with microbes
Next stop?
Time to learn all about
bacteria and archaea
visualize colorless bacteria under the microscope
"heat fix" bacteria to slide during smear preparation
many stains to choose from, depends on what you are looking for
Inoculation
Incubation
Isolation
Inspection
Information gathering
Identification
Magnification-make things
bigger
Resolution-ability to see
detail and distinguish objects
as separate
Types of Microscopes
Media
provide microbes with
nutrients in an artificial
medium
more than 500 different
types
microbes introduced by loop,
needle, pipette, or swab
fig. 3.1, pg. 60
pg. 61
fig. 3.17, pg. 76
fig. 3.18, pg. 77
fig. 3.19, pg. 77
Isolation Methods
How isolation works
Streak Plate
best for isolated colonies
that can be used for
for subcultures
bacteria on surface of agar
Pour plate
best for counting colonies
bacteria grow all throughout
the agar
Spread plate
best if plate has antibiotic
selection or if you want to
plate large amount of broth
culture
Pure
Mixed
Contaminated
Parts of the microscope
Only need to know for lab
virtual image appears
upside down and inverted
Magnification occurs twice
Objective lens
Ocular lens
Total magnification equals
power of objective X
power of ocular
Comparison of Microscopes
Resolution is dependent on
numerical aperature of lens
and wavelength.
The wavelength of visible light
waves limits ability of light
microscope.
Shorter wavelengths resolve better!
Resolution can be improved by
adding oil that has same refractive
index as glass. Oil immersion lens
sits in oil and prevents loss of light
to the atmosphere.
Maximum resolution of
oil immersion lens is 0.2 micrometers.
Brightfield
Dark field
Phase contrast
Fluorescent
Confocal
DIC
Electron microscopes magnify
many times more than light
microscopes because electrons
can travel in wavelengths 100,000X
shorter than visible light waves.
Transmission electron microscope (TEM)
Scanning electron
microscope (SEM)
fig. 3.3, pg. 62
fig. 3.4, pg. 63
fig. 3.5, pg. 63
fig. 3.6, pg. 64
fig. 3.7, pg. 64
pg. 65
fig. 3.8, pg. 66
fig. 3.9, pg. 66
fig. 3.10, pg. 67
fig. 3.11, pg. 67
fig. 3.12, pg. 67
fig. 3.13, pg. 68
fig. 3.14, pg. 69
opposite charges
attract and dye
penetrates into cell
like charges repel
and dye stays outside
of cell. background is
colored, while cell is
clear
Blood agar
Chocolate agar
fig. 3.15, pg. 71
pg. 71
fig. 3.16, pg. 72
pg. 79
fig. 3.21, pg. 79
fig. 3.22, pg. 80
Liquid media
urea broth
presence-absence
broth
Semisolid media
sulfur indole
motility media
pg. 81
Enriched Media
fig. 3.25, pg. 82
fig. 3.24, pg. 82
Media that are
selective and differential
fig. 3.26, pg. 83
mannitol salt agar
MacConkey agar
fig. 3.28, pg. 85
fig. 3.27, pg. 84
differentiate multiple characteristics
triple sugar
iron agar
differentiate between common microbes that
cause UTIs
carbohydrate
fermentation
broth
Full transcript