Part one: Spots

1) Spots of Microbiological
Culture Media and
Biochemical Reactions
 1-A-Ordinary liquid media
Name: Nutrient Broth
(Clear yellowish fluid)
Type : Ordinary liquid media
Result:
Control +ve Growth
(clear) (turbid)
 1-B- Ordinary solid media
Name : Nutrient Agar plate
Type: Ordinary Solid Media
Result:
Control +ve (growth)
Control +ve (growth)
 2- Enriched media
Name : Blood agar
Type : Enriched Media
Result :
1) control
2) β- hemolysis on blood agar
(clear zone around growth)
3) α- hemolysis on blood agar
(greenish discoloration)

4) γ- non-hemolysis on blood agar (growth only)

Name :Chocolate agar
Type : Enriched Media
Result:

control
3- Selective and Differential media
1. Mannitol salt agar (MSA)
2. Macconkey agar medium
3. Eosin-methylene blue medium(EMB)
Name : Mannitol salt agar (MSA)
Type: Selective and Differential media
Result:

1) control
2) Mannitol Fermenter

3) Mannitol- Non fermenter

Name : MacConkey agar medium
Type: Selective and Differential media
Result:
1) control
2) Lactose Fermenter( Rose pink colonies)

3) Lactose Non fermenter ( pale yellow colonies)

Name: Eosin-methylene blue agar(EMB agar)
Type : Selective and Differential media
Result:
1) Control

2) Lactose Fermenter ( dark/blackish colonies)

3) Extreme lactose fermenter (dark colonies with
green metallic sheen of E.coli)

4) Lactose Non fermenter (colorless colonies)

4- Differential and indicator media

1- Simmons citrate agar

2-triple sugar iron (TSI)agar
Name : Simmons citrate agar
Type: Differential and indicator media
Result :
1) Control is Green.
2) +ve growth is blue.
Name : triple sugar iron (TSI)agar
Type: Differential and indicator media
Result:
Results of TSI
 Biochemical Reactions
IMViC test
O/F test (Oxidation/Fermentation test)
Motility test
Nitrate reduction test
Urease test
Catalase test
Coagulase test
 IMViC test

A) Indole test
B) Methyl Red test
C) Voges Proskauer test
D) Citrate Utilization test
Name: Indole test.
Result:
Indole +ve Indole -ve
Name: Methyl red test
Result:

Methyl red Methyl red

+ve -ve
Name: Voges Proskauer test
Result:

Voges Proskauer Voges Proskauer

+ve -ve
Name: citrate utilization test
Result:

Citrate utilization Citrate utilization

+ve -ve
Results of IMViC test
Name : O/F test (Oxidation/Fermentation test)
Result:

O+/F+ O+/F-
Name : Motility test
Result:

Motile Non-motile
Name : Nitrate reduction test
Result:

+ve (red col.) -ve (yellowish)

Name : Urease test
Result:

+ve (pink col.) -ve (yellow)

Name: Catalase test.
Result:

+ve (effervescence) -ve

Name: Coagulase test
Result:
Coagulase +ve (clotting) Coagulase -ve ( no clotting)
Control (C): liquid Control (C): liquid
Test (T): solid Test (T): liquid
 2) Spots of Immunology
(Agglutination Reactions)
 1) The White Blood Cells

Name : Microscopical Examination of Neutrophile .

Type: Granulocyte.
Description: multilobed nucleus (3-5 lobes) connected
with filaments , blue, with numerous fine granules.
Name : Microscopical Examination of Neutrophile .
Type: Granulocyte.
Description: bilobed nucleus ,No connected filaments,
stained blue with coarse red granules.
Name : Microscopical Examination of Neutrophile .
Type: Agranulocyte.
Description: The nucleus is rounded, occupies most of
the cell and stained blue.
Name : Microscopical Examination of Neutrophile .
Type: Agranulocyte.
Description: The nucleus is kidney-shaped and occupies
less volume of the cell and stained blue.
 2) Agglutination Reactions
Name: Blood Grouping.
Type: Slide Agglutination Reaction
Result: Determination blood group/type.
Name: Widal test.
Type: Tube agglutination reaction
Result (interpretation): titer = 40
Uses of test: used for diagnosis of typhoid fever
Name: Pregnancy test.
Type: Latex agglutination reactions
Result:
 The test is considered +ve if you find 2 red lines
 The test is considered -ve if you find a single red line only
Uses: detection of the pregnancy.
3) Spots of Staining
1)Parts of microscope and their
functions
 Arm: Supports the tube and connects it to the base
 Base: The bottom of the microscope, used for
support
 Eyepiece (ocular lens): Where you look to see the
image of your specimen.
 Body tube: Connects the eyepiece to the objective
lenses.
 Revolving Nosepiece: holds objective lenses and
can be rotated to easily change the magnification
power.
 Objective lenses: used to increase the
magnification of the specimen.
 - scaning (4X) - Low power (10X)
 - high power (40X) - Oil immersion (90X or
100X
 Stage: The flat platform where you place your slides.
 Aperture: the hole in the stage through which the transmitted
light reaches the object to be examined.
 Stage Clips: hold the slide in place
 Condenser: used to collect and focus the light from the
light source on the specimen. It is located under the stage
often in conjunction with an iris diaphragm.
 Iris Diaphragm: controls the light going thorough the
aperture.
 Coarse Adjustment Knob: large, round knob on the side
of the microscope used for bringing the object into
approximate focus.
 Fine Adjustment Knob: small, round knob on the side of
the microscope used to bring the object into perfect focus
 Mirror/Light source: used to reflect light to the specimen
Definitions
 Magnification: how much an image is enlarged
 Magnification power of microscope = magnification of
ocular lens x magnification of objective lens
 Resolution: the amount of details that you can see of
an image. resolution = WL / 2NA
1- wavelength (WL) of light source resolution
so, the electron microscope has higher resolution
than the compound microscope
2- numerical aperture (NA) resolution
NA: the amount of light which enters the objective.
The oil immersion objective has the highest NA, so
produces the highest resolution
3- Refractive index:
When the light passes from the light source through
the slide and into the oil immersion lens, it will be
refracted as it passes through glass (slide) and air
(between the slide and the lens). The image can not
be seen with a good resolution
Thus, a drop of cedar wood oil is put on the film as
it has the same refractive index of glass. Thus, it
prevents light refraction
 2)Simple Staining
A) By using Crystal Violet

1)Name: simple stain by using crystal violet.

Description: oval , single , violet.

2)Name: simple stain by using crystal violet.

Description: Rods, single , violet.
3)Name: simple stain by using crystal
violet.
Description: Cocci , in bunches , violet.

4)Name: simple stain by using crystal

violet.
Description: Bacilli , in chains , violet.
B) By using Methylene Blue
1)Name: simple stain by using methylene blue.
Description: oval , single , blue.

2)Name: simple stain by using methylene

blue
Description: Rods, single , blue.
3)Name: simple stain by using methylene blue
Description: Cocci , in bunches , blue.

4)Name: simple stain by using methylene

blue
Description: Bacilli , in chains , blue.
C) By using Safranin

1)Name: simple stain by using Safranin

Description: oval , single , red.

2)Name: simple stain by using Safranin

Description: Rods, single , red.

3)Name: simple stain by using Safranin
Description: Cocci , in bunches , Red.

4)Name: simple stain by using Safranin

Description: Bacilli , in chains , Red.
 3) Negative staining
1)Name: negative staining by nigrosin
Description: colorless bacilli in chain on dark background.

2)Name: negative staining by nigrosin

Description: colorless oval , single on
dark background.
 4) Acid Fast Staining
Name: Acid fast staining of Mycobacterium Pheli
Description: long rods, single, dark red.
Mycobacterium (B)>>>dark red ,long rods with cocci
(A)>>blue
 5) Endospore staining
Name : Endospore staining of bacilli spp.
Description: vegetative bacilli in chains(red)
contain green colored endospore.
 Part two:
Gram stain Mixture