Things to Do

1
Things to Do

• Scope lecture
• Streak pour plate results
• Staining

2
Microscopy
3
Scope Image Comparison
4
Comparison of Scopes
Used with permission
CDC, used with permission
Darkfield
Brightfield
Wikipedia Commons, Used with permission
Phase-Contrast
5
Comparison of Microscopes
6
Fluorescent Microscopy
7
Immunofluorescent staining
8
Immunofluorescent staining of influenza virus
9
Immunofluorescent staining
10
Advantage of using Fluorescence

• Without stain

• With Stain

11
(No Transcript)
12
Streak plate
Review common mistakes
13
Pour Plate
14
Plate Counts
Used with permission
15
(No Transcript)
16
Cryptococcus neoformans in CSF
WebPath, used with permission
17
Capsule Stain
18
Capsule stains
Used with permission
19
Flagella Stain
KM
20
Flagella Stain
KM
21
Mechanism of the Acid-Fast Stain
Non acid-fast
Acid-Fast

• Before staining
• Steam with
• Carbolfuchsin
• Acid alcohol
• Decolorization
• Counterstain with
• Methylene blue

22
The Ziehl-Neesen Acid-Fast Stain
What do you observe? What do you conclude?
23
AFB
WebPath, used with permission
24
AFB via fluroscence
WebPath, used with permission
25
Mechanism of the endospore stain
Makes endspores
Lacks endospores

• Before staining
• Steam with
• Malachite green
• Wash with water
• Counterstain with
• safranin

26
Bacillus endospores
24 hr
KM
27
Bacillus endospores
3 Days
Used with permission
28
Bacillus endospores

• 5 Days

• 7 Days

Used with permission
29
(No Transcript)
30
Bacillus
Unstained endospores
KM
31
Clostridium tetani
32
Clostridium tetani
CDC, used with permission
33
Bacillus endospores
KM
1 Week
34
Immunofluorescent staining
35
Immunofluorescent staining
36
Darkfield
37
Capsule Stain
KM
38
Bacillus endospores
3 Days