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A diagram of serial dilution. A large beaker on the left contains a dark solution. 1 ml is moved from this beaker to a tube containing 9 ml of broth. This tube has a dilution of 1:10 and is lighter in color than the original beaker. A sample of 0.1 ml from this tube is put on an agar plate; the colonies are too numerous to count. 1 ml is taken out of this tube and placed in a tube containing 9 ml of broth. This tube now has a dilution of 1:100 from the original beaker and is even lighter in color. 0.1 ml is plated on an a agar plate and the colonies are still too numerous to count. 1 ml is taken from this tube and placed in another tube containing 9 ml broth. This is now a dilution of 1:1000 from the original beaker and the tube is lighter than the last. 0.1 ml is taken out of this tube and placed on an agar plate; there are 389 colonies. 1 ml is taken out of this tube and placed in another tube containing 9 ml broth. This is now a dilution of 1:10,000 from the original beaker and this tube is even lighter than the last. 0.1 ml is taken out of this tube and placed on an agar plate; there are 50 colonies. 1 ml is taken out of this tube and placed in a tube containing 9 ml of broth. This is a dilution of 1:100,000 from the original beaker and this is the lightest tube of all. 0.1 ml is taken from this tube and placed on an agar plate; there are 2 colonies.
Serial dilution involves diluting a fixed volume of cells mixed with dilution solution using the previous dilution as an inoculum. The result is dilution of the original culture by an exponentially growing factor. (credit: modification of work by “Leberechtc”/Wikimedia Commons)

The dilution factor is used to calculate the number of cells in the original cell culture. In our example, an average of 50 colonies was counted on the plates obtained from the 1:10,000 dilution. Because only 0.1 mL of suspension was pipetted on the plate, the multiplier required to reconstitute the original concentration is 10 × 10,000. The number of CFU per mL is equal to 50 × 100 × 10,000 = 5,000,000. The number of bacteria in the culture is estimated as 5 million cells/mL. The colony count obtained from the 1:1000 dilution was 389, well below the expected 500 for a 10-fold difference in dilutions. This highlights the issue of inaccuracy when colony counts are greater than 300 and more than one bacterial cell grows into a single colony.

A diagram of the pour plate method. Step 1 – the bacterial sample is mixed with warm agar (45-50° C). Step 2 – the sample is poured onto a sterile plate. Step 3 – the sample is swirled to mix and allowed to solidify. Step 4 – the plate is incubated until bacterial colonies grow.
In the pour plate method of cell counting, the sample is mixed in liquid warm agar (45–50 °C) poured into a sterile Petri dish and further mixed by swirling. This process is repeated for each serial dilution prepared. The resulting colonies are counted and provide an estimate of the number of cells in the original volume sampled.
A diagram of the spread plate method. Step 1 – a sample (0.1 ml) from a bacterial dilution is poured onto a solid medium. Step 2 – the sample is spread evenly over the surface. Step 3 – the plate is incubated until bacterial colonies grow on the surface of the medium.
In the spread plate method of cell counting, the sample is poured onto solid agar and then spread using a sterile spreader. This process is repeated for each serial dilution prepared. The resulting colonies are counted and provide an estimate of the number of cells in the original volume samples.

A very dilute sample—drinking water, for example—may not contain enough organisms to use either of the plate count methods described. In such cases, the original sample must be concentrated rather than diluted before plating. This can be accomplished using a modification of the plate count technique called the membrane filtration technique . Known volumes are vacuum-filtered aseptically through a membrane with a pore size small enough to trap microorganisms. The membrane is transferred to a Petri plate containing an appropriate growth medium. Colonies are counted after incubation. Calculation of the cell density is made by dividing the cell count by the volume of filtered liquid.

The most probable number

The number of microorganisms in dilute samples is usually too low to be detected by the plate count methods described thus far. For these specimens, microbiologists routinely use the most probable number (MPN) method , a statistical procedure for estimating of the number of viable microorganisms in a sample. Often used for water and food samples, the MPN method evaluates detectable growth by observing changes in turbidity or color due to metabolic activity.

Questions & Answers

what is fermentation example ?
Sonal Reply
is proceess in which an agent couses of an oganic substances breakdown into simpler substance,especially in aneorobic breakdown of suger into alcohol.
Okashat
is it better to study microbiology and then medicine it makes no difference to go directly to medicine?
Jessee Reply
Dray's mathdme cell wall konse color k hote he
Jinal Reply
what is dray's mathdme cell wall
Prabhat
I confused. please help me
Karen
just confused
Raj
l don't understand it please explain it for me.
Karen Reply
epitopes are present on the surface of
Rohit Reply
at the tip of variable region on the antibody...where antigen and antibody binding sites combine...
Fiza
The term that is used refer to moving microbes under a microscope are referred to as?
Lee Reply
Members of the genus Neisseria cause which of the folowing human diseases?
Farah Reply
genital infections
Kamaluddeen
gonorrhoea
sandip
gonorrhoea
Jessee
4. Which of the following specimens should not be refrigerated? a. Urine b. Urogenital swab
Zahraa Reply
urine
Muuse
urine
Agatha
Urine
Tean
urine
Yasser
Urine
Ebtehal
urine
anamika
urine
Puja
urine
Inemesit
urine
Samuel
Urine
Muhammad
Details about McConkey agar
Muhammad
urine
SK
urine
what is bacteria
anamika Reply
a member of large number of unicellular microorganism which have cell wall but lack of cell organelles an oranised nucleus including somewhat can cause disease
Sukhdeep
Bacteria are usually composed of one cell onl to that are neither plants nor animals, microscopic, that may cause diseases or may be beneficial(in gut)... it depends upon their weapons. Nearly all animal life is dependent on Bacteria for their survival
Fiza
thanks
anamika
what factor make bacteria colony large and how could we sterlise it in large scale
fatty
nutrient concentration temp gaseous conc ph ion or salt concentration mositure condition factors contribute to make large colony. by autoclaving we will sterilize bactetia
Sukhdeep
Colony is actually visible growth of Bacteria that is as a result of suitable environment for growth i.e optimal conditions for growth, temperature, moisture etc. there're many methods to get rid of bacteria. If We stop giving them optimal conditions for living Bacteria will die soon .
Fiza
what's the difference between an antigen and a pathogen?
Pathogens are organisms that cause disease in other organisms whereas Antigen is a part of a pathogen that triggers the immune response..
Rajat
so it is the antigen that dendritic cells present to the T cells and not the pathogen itself?
no no antigen are the west product or part of the pathogen. in such case bacteria it self fight with over immune response & in another case bacteria release antigens
vasava
& other antigen like pollan grain, dust particles etc.....
vasava
pathogen are microbes that can infect the body and causw illness....antigens are the part of pathogens that alert the body to an infection
Sukhdeep
antigen is a part of blood and pathogen is foreign particle which causes diseases
Yogyata
antigen could be non microorganism.... where as pathogen is mixroorganism
tadesse
Thanks
Karen
a pathogen is a disease causing organism while an antigen is a protein in the white blood cells which combats pathogens.
Jessee
what type of widal test
sobhit Reply
this test determine for typhoid in this test if H,O antigen are present that indicate the positive test bac. are salmonella typhy
vasava
what h.o denotes
Iqra
o: body of bacteria, h: flagellate
Explain Mould
Chinenye Reply
Explain mycoses and it's classification
Chinenye
why do we have hiccups?
Manisha Reply
shakey diaphragm
Curlisse
The antibody binding site is formed primarily by:
Asalla Reply
How many types of MICROORGANISMS do we have?
Hope Reply
Hello friends
Hope
microorganisms are divided into seven type Bacteria archaea protozoa algae fungi virus and multicellular animal parasites
Raj

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Source:  OpenStax, Microbiology. OpenStax CNX. Nov 01, 2016 Download for free at http://cnx.org/content/col12087/1.4
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