An agar plate is a sterile Petri dish that contains a

growth medium (typically agar plus nutrients) used to culture

microorganisms or small plants like the moss
Physcomitrella patens.
Selective growth compounds may also be added to the media,
such as antibiotics.
Individual microorganisms placed on the plate will grow into
individual colonies, each a clone genetically identical to the
individual ancestor organism (except for the low, unavoidable
rate of mutation).
Thus, the plate can be used either to estimate the
concentration of organisms in a liquid culture or a suitable
dilution of that culture, using a colony counter, or to generate
genetically pure cultures from a mixed culture of genetically
different organisms, using a technique known as streaking.

In this technique, a drop of the culture on the

end of a thin, sterile loop of wire is "streaked"
across the surface of the agar leaving
organisms behind, a higher number at the
beginning of the streak and a lower number
at the end.
At some point during a successful "streak",
the number of organisms deposited will be
such that distinct individual colonies will grow
in that area which may be removed for
further culturing, using another sterile loop. [1]

A Petri dish is a shallow glass or plastic

cylindrical lidded dish that biologists use to culture
cells. It was named after German bacteriologist
Julius Petri, who invented it when working as an
assistant to Robert Koch. Glass Petri dishes can be
re-used by sterilization (for example, dry heating
in a hot air oven at 160 C for one hour); plastic
Petri dishes must be disposed of after one use.
For microbiology, agar plates are very frequently
used. The dish is partially filled with warm liquid
agar along with a particular mix of nutrients, salts
and amino acids and, optionally, antibiotics. After
the agar solidifies, the dish is ready to receive a
microbe-laden sample.

Other Petri dish uses do not involve agar; for instance, cell culture.
Modern Petri dishes often have rings on the lids and bases which
allow them to be stacked so that they do not slide off one another.
Multiple dishes can also be incorporated into one plastic container to
create what is called a "multi-well plate".
As well as making agar plates, empty Petri dishes may be used to
observe plant germination or small animal behaviour, or for other
day-to-day laboratory practices such as drying fluids in an oven and
carrying and storing samples.

A growth medium or culture medium is

a liquid or gel designed to support the
growth of microorganisms or cells.
There are different types of media for
growing different types of cells.
There are two major types of growth media:

those used for cell culture, which use specific

cell types derived from plants or animals,
and microbiological culture, which are used for
growing microorganisms, such as bacteria or
yeast.

The most common growth media for

microorganisms are nutrient broths and
agar plates; specialized media are sometimes
required for microorganism and cell culture
growth.[1]
Some organisms, termed fastidious
organisms, require specialized environments
due to complex nutritional requirements.
Viruses, for example, are obligatory
intracellular parasites and require a growth
medium composed of living cells.

The most common growth media for

microorganisms are nutrient broths
(liquid nutrient medium) or
Luria Bertani medium (LB medium or
Lysogeny Broth).
Liquid media are often mixed with agar
and poured into petri dishes to solidify.
These agar plates provide a solid medium
on which microbes may be cultured.
Bacteria grown in liquid cultures often
form colloidal suspensions.

The differences between growth media used

for cell culture and those used for
microbiological culture are because cells
derived from whole organisms and grown in
culture often cannot grow without the
addition of, for instance, hormones or
growth factors which usually occur in vivo.[3]
In the case of animal cells, this difficulty is
often addressed by the addition of
blood serum to the medium.

In the case of microorganisms, there are no

such limitations, as they are often
unicellular organisms.
One other major difference is that animal cells
in culture are often grown on a flat surface to
which they attach, and the medium is
provided in a liquid form, which covers the
cells.
In contrast, bacteria such as Escherichia coli
may be grown on solid media or in liquid
media.

An important distinction between

growth media types is that of defined
versus undefined media.

A defined medium will have known quantities

of all ingredients.

For microorganisms, they consist of providing

trace elements and vitamins required by the
microbe and especially a defined
carbon source and nitrogen source.

Glucose or glycerol are often used as carbon

sources, and ammonium salts or nitrates as
inorganic nitrogen sources).

An undefined medium has some complex

ingredients, such as yeast extract or casein
hydrolysate, which consist of a mixture of
many, many chemical species in unknown
proportions.

Undefined media are sometimes chosen

based on price and sometimes by necessity
- some microorganisms have never been
cultured on defined media.

A good example of a growth medium is the

wort used to make beer.
The wort contains all the nutrients
required for yeast growth, and under
anaerobic conditions, alcohol is produced.
When the fermentation process is
complete, the combination of medium and
dormant microbes, now beer, is ready for
consumption

An undefined medium (also known as a basal or complex

medium) is a medium that contains:
a carbon source such as glucose for bacterial growth
water
various salts needed for bacterial growth
a source of amino acids and nitrogen (e.g., beef, yeast
extract)
This is an undefined medium because the amino acid
source contains a variety of compounds with the exact
composition being unknown.
Nutrient media contain all the elements that most
bacteria need for growth and are non-selective, so they
are used for the general cultivation and maintenance of
bacteria kept in laboratory culture collections.

Defined media (also known as chemically

defined media)

all the chemicals used are known

does not contain any animal, yeast, plant tissue.

Differential medium
some sort of indicator, typically a dye, is
added, that allows for the differentiation of
particular chemical reactions occurring
during growth.

Minimal media

Minimal media are those that contain the

minimum nutrients possible for colony
growth, generally without the presence of
amino acids, and are often used by
microbiologists and geneticists to grow "wild
type" microorganisms.

Minimal media can also be used to select for

or against recombinants or exconjugants.

Minimal medium typically contains:

a carbon source for bacterial growth, which

may be a sugar such as glucose, or a less
energy-rich source like succinate
various salts, which may vary among
bacteria species and growing conditions;
these generally provide essential elements
such as magnesium, nitrogen, phosphorus,
and sulfur to allow the bacteria to synthesize
protein and nucleic acid
water

Supplementary minimal media

are a type of minimal media that also

contains a single selected agent, usually
an amino acid or a sugar.
This supplementation allows for the
culturing of specific lines of auxotrophic
recombinants.

Bile esculin agar (BEA)

BEA is used for the isolation of Enterococci as well
as Group D Streptococci
Cysteine Lactose Electrolyte Deficient agar (CLED)
CLED agar is used to isolate and differentiate
urinary tract bacteria, since it inhibits Proteus
species swarming and can differentiate between
lactose fermenters and non-fermenters.
Hektoen enteric agar (HEA)
HE agar is designed to isolate and recover fecal
bacteria belonging to the Enterobacteriaceae family.
HE is particularly useful in isolating Salmonella and
Shigella.

Lysogeny Broth (LB)

MacConkey agar (MAC)
A selective and differential media used to
differentiate between Gram negative bacteria while
inhibiting the growth of Gram positive bacteria. The
addition of bile salts and crystal violet to the agar
inhibits the growth of most Gram positive bacteria,
making MacConkey agar selective. Lactose and
neutral red are added to differentiate the lactose
fermenters, which form pink colonies, from lactose
nonfermenters that form clear colonies. An
alternative media, eosin methylene blue (EMB)
serves a similar purpose.

Mannitol salt agar (MSA)

MSA is also a selective and differential media.
Mannitol is the differential part, it indicates
organisms that ferment mannitol. If mannitol
fermentation is occurring, lactic acid will be
produced, and the pH will drop causing the
MSA plate to turn yellow. The salt portion is
selective for halophiles; organisms that
cannot withstand a high salt content will be
unable to grow well. Mueller-Hinton agar
(MHA)

MHA contains beef infusion, peptone, and starch

used primarily for antibiotic susceptibility
testing. It can be in a form of blood agar.
Fungi growing in axenic culture (ascomycetes)
Nutrient agar
Nutrient agar is usually used for growth of nonfastidious organisms and observation of
pigment production. It is safe to use in school
science laboratories because it does not
selectively grow pathogenic bacteria.

Selective media are used for the growth of only

select microorganisms.
For example, if a microorganism is resistant to a
certain antibiotic, such as ampicillin or
tetracycline, then that antibiotic can be added to
the medium in order to prevent other cells, which
do not possess the resistance, from growing.
Media lacking an amino acid such as proline in
conjunction with E. coli unable to synthesize it
were commonly used by geneticists before the
emergence of genomics to map bacterial
chromosomes.

Selective growth media are also used in

cell culture to ensure the survival or
proliferation of cells with certain properties,
such as antibiotic resistance or the ability to
synthesize a certain metabolite.

Normally, the presence of a specific gene or

an allele of a gene confers upon the cell the
ability to grow in the selective medium. In
such cases, the gene is termed a marker.

Selective growth media for eukaryotic cells

commonly contain neomycin to select cells
that have been successfully transfected
with a plasmid carrying the neomycin
resistance gene as a marker.

Gancyclovir is an exception to the rule as it

is used to specifically kill cells that carry its
respective marker, the Herpes simplex virus
thymidine kinase (HSV TK).

Some examples of selective media include:

eosin-methylen blue agar (EMB) that contains
methylene blue toxic to Gram-positive
bacteria, allowing only the growth of
Gram negative bacteria
YM (yeast and mold) which has a low pH,
deterring bacterial growth
blood agar (used in strep tests), which contains
beef heart blood that becomes transparent in
the presence of hemolytic Streptococcus

MacConkey agar for Gram-negative

bacteria
Hektoen Enteric (HE) which is selective for
Gram-negative bacteria
Mannitol Salt Agar (MSA) which is
selective for Gram-positive bacteria and
differential for mannitol
Terrific Broth (TB) is used with glycerol in
cultivating recombinant strains of
Escherichia coli.

xylose lysine desoxyscholate (XLD),

which is selective for Gram-negative
bacteria
Buffered charcoal yeast extract agar,
which is selective for certain gramnegative bacteria, especially
Legionella pneumophila

Four types of agar plates

demonstrating differential growth
depending on bacterial metabolism.

Differential media or indicator media

distinguish one microorganism type from
another growing on the same media.
This type of media uses the biochemical
characteristics of a microorganism growing in
the presence of specific nutrients or
indicators (such as neutral red, phenol red,
eosin y, or methylene blue) added to the
medium to visibly indicate the defining
characteristics of a microorganism.
This type of media is used for the detection of
microorganisms and by molecular biologists
to detect recombinant strains of bacteria.

Examples of differential media include:

Eosin methylene blue (EMB), which is
differential for lactose and sucrose
fermentation
MacConkey (MCK), which is differential for
lactose fermentation
Mannitol Salt Agar (MSA), which is
differential for mannitol fermentation
X-gal plates, which are differential for
lac operon mutants

These are used for the temporary

storage of specimens being transported
to the laboratory for cultivation.
Such media ideally maintain the
viability of all organisms in the
specimen without altering their
concentration.

Transport media typically contain only

buffers and salt.
The lack of carbon, nitrogen, and
organic growth factors prevents
microbial multiplication.
Transport media used in the isolation of
anaerobes must be free of molecular
oxygen. Example:stuart transport
medium.

Enriched media contain the nutrients

required to support the growth of a
wide variety of organisms, including
some of the more fastidious ones.
They are commonly used to harvest as
many different types of microbes as are
present in the specimen.

Blood agar is an enriched medium in

which nutritionally rich whole blood
supplements the basic nutrients.
Chocolate agar is enriched with heattreated blood (40-45C), which turns
brown and gives the medium the color
for which it is named.

MacConkey (also McConkey) agar is a

culture medium designed to grow
Gram-negative bacteria and stain them for
lactose fermentation.
Contents

It contains bile salts (to inhibit most Gram-positive

bacteria, except Enterococcus and some species
of Staphylococcus ), crystal violet dye (which also
inhibits certain Gram-positive bacteria),
neutral red dye (which stains microbes fermenting
lactose), lactose and peptone.

A MacConkey agar plate with an

active bacterial culture.

Mac Conkey agar with lactose and

non-lactose fermenters

Alfred Theodore MacConkey developed

it while working as a bacteriologist for
the Royal Commission on Sewage
Disposal.

Acting as a visual pH indicator, the agar

distinguishes those Gram-negative bacteria
that can ferment the sugar lactose (Lac+)
from those that cannot (Lac-).

By utilizing the lactose available in the

medium, Lac+ bacteria such as
Escherichia coli, Enterobacter and Klebsiella
will produce acid, which lowers the pH of the
agar below 6.8 and results in the
appearance of red/pink colonies.

Non-Lactose fermenting bacteria such

as Salmonella, Proteus species and
Shigella cannot utilize lactose, and will
use peptone instead.

This forms ammonia, which raises the

pH of the agar, and leads to the
formation of white/colorless colonies.

Thus, by selecting for Gram-negative

bacteria and differentiating between enteric
pathogens such as E. coli and Salmonella,
the MacConkey agar is readily implemented
in the clinical diagnosis of diarrhea.

This medium is also known as an "indicator

medium" and a "low selective medium".
Absence of electrolytes serves to inhibit
swarming by Proteus species

Cetrimide agar is a type of agar used for the

selective isolation of the gram-negative bacteria,
Pseudomonas aeruginosa.
As the name suggests, it contains cetrimide, which
is the selective agent against alternate microbial
flora.
Cetrimide also inhances the production of
Pseudomonas pigments such as pyocyanin and
fluorescein, which show a characteristic bluegreen and yellow-green colour, respectively.

[3] [4]

Cetrimide agar is widely used in the examination

of cosmetics, pharmaceuticals and clinical
specimens to test for the presence of
Pseudomonas aeruginosa

Xylose lysine deoxycholate agar (XLD

agar) is a selective growth medium used in
the isolation of Salmonella and Shigella
species from clinical samples and from food.
It has a pH of approximately 7.4, leaving it
with a bright pink or red appearance due to
the indicator phenol red.
Sugar fermentation lowers the pH and the
phenol red indicator registers this by
changing to yellow.
.

Most gut bacteria, including Salmonella, can ferment

the sugar xylose to produce acid; Shigella colonies
cannot do this and therefore remain red.

After exhausting the xylose supply Salmonella

colonies will decarboxylate lysine, increasing the pH
once again to alkaline and mimicking the red Shigella
colonies.

Salmonellae metabolise thiosulfate to produce

hydrogen sulfide, which leads to the formation of
colonies with black centers and allows them to be
differentiated from the similarly coloured Shigella
colonies

Other Enterobacteria such as E. coli will

ferment the lactose and sucrose present in
the medium to an extent that will prevent
pH reversion by decarboxylation and
acidify the medium turning it yellow.
Salmonella species: red colonies, some
with black centers.
Shigella species: red colonies.
Coliforms: yellow to orange colonies.
Pseudomonas aeruginosa: pink, flat, rough
colonies

Yeast extract

3g/l

L-Lysine

5g/l

Xylose

3.75g/l

Lactose

7.5g/l

Sucrose

7.5g/l

Sodium deoxycholate

1g/l

Sodium chloride

5g/l

Sodium thiosulfate

6.8g/l

Ferric ammonium citrate

0.8mg/l

Phenol red

0.08g/l

Agar

12.5g/

nz agar
nz agar allows more rapid bacteriological
diagnosis as Salmonella and Shigella colonies can
be clearly and reliably differentiated from other
Enterobacteriaceae. The yields of Salmonella from
stool samples obtained, when using this medium,
are higher than those obtained with LEIFSON Agar
or SalmonellaShigella agar (SSA).
Phenylethyl alcohol agar (PEA)
PEA selects for Staphylococcus species while
inhibiting Gram-negative bacilli (e.g. Escherichia coli
, Shigella, Proteus, etc.). MSA is selective for
Staphylococcus PEA is selective for gram positive
bacteria

R2A Agar (R2A)

A non-specific agar that imitates the medium of
water. Used for water analysis.
Tryptic (Trypticase) Soy Agar (TSA)
TSA is a general purpose media produced via
enzymatic digestion of soybean meal and casein
; TSA is frequently the base media of other agar
plate type, i.e. blood agar plates (BAP) are made
by enriching TSA plates with blood (see above).
TSA plates support growth of many semifastidious bacteria, including some species of
Brucella, Corynebacterium, Listeria, Neisseria,
and Vibrio.

Xylose-Lysine-Deoxycholate agar (XLD)

XLD is used for the culture of stool samples, and
contains two indicators. It is formulated to
inhibit Gram-positive bacteria, while the growth
of Gram-negative bacilli is encouraged. The
colonies of lactose fermenters appear yellow. It
is also used to culture possible Salmonella that
may be present in a food sample. Salmonella
colonies will show a black halo on XLD.
Cetrimide agar
Cetrimide agar is a type of agar used for the
selective isolation of the gram-negative
bacteria, Pseudomonas aeruginosa

Sabouraud agar
Sabouraud agar is used to culture fungi and
has a low pH that inhibits the growth of most
bacteria; also contains the antibiotic
gentamicin to specifically inhibit the growth of
Gram-negative bacteria. Hay Infusion agar
Specific for the culturing of slime moulds
(though not technically fungi). Potato dextrose
agar
PDA is used to culture of certain types of fungi.