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Streak Plate Method - Principle, Types, Methods, Uses
Streak Plate Method - Principle, Types, Methods, Uses
Streak Plate Method - Principle, Types, Methods, Uses
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Home » Basic Microbiology » Streak Plate Method- Principle, Types, Methods, Uses
Streak literally means “a long, thin line”: and the streak plate method is a microbiological
culture technique where a sample is spread in a petri dish in the form of a long, thin line over
the surface of solid media.
Table of Contents
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This method dilutes the bacterial load, over the surface of agar medium, successively as
streaking proceeds, and ultimately only a few bacterial cells will be inoculated at the end
giving well-isolated colonies in the final streaks. Thus, this method mechanically isolated the
bacteria from a mixed population of either the same or different species. After inoculation,
the same types of colonies are seen in the terminal streaks if the specimen contained single
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species, whereas, different types of colonies may be seen if the specimen contained different
species.
This is a very old method used in microbiology since the time of Robert Koch. This method
was first devised and used by Loeffler and Gaffky in Koch’s laboratory to serially dilute
bacteria over agar surface and obtain well-isolated colonies. Since that time it is used as a
very important tool in bacteriology.
It is a very simple and reliable aseptic technique that uses tools like cotton swabs, wooden or
plastic, metal sticks and toothpicks, or inoculating loop to dilute and spread the specimen
over the surface of pre-sterilized specific solid culture media. The specimen used can be
either suspension or colonies from the agar surface. Well isolated colonies can be obtained
from successfully performed streaking which allows describing the colony character of the
organism on that specific culture media and condition.
Astronomers Track Matter Traveling Faster Than Light, Seemingly Breaking th…
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1. Quadrant Streaking
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It is the most commonly used and the most preferred method where four equal-sized
sections of the agar plate are streaked. It is also referred to as the “four-quadrant streak” or
“four sectors” or “four-way streak” method.
In this method, each plate is divided into four equal sectors and each adjacent sector is
streaked sequentially. The sector which is streaked first is called the first sector or the first
quadrant, and it has the highest concentration of inoculum. Gradually the second, third, and
fourth quadrants will have diluted inoculum. By the time the fourth quadrant is streaked, the
inoculum is highly diluted giving rise to isolated colonies following the incubation.
Mostly, a discontinuous fashion of streaking is followed where the loop is sterilized at the end
of each quadrant prior to streaking over the next quadrant. However, if the bacterial load is
too small (or highly diluted), continuous fashion can also be used. In the latter, the loop needs
not be sterilized at the end of every quadrant.
Although being the most popular method, it limits us to use only one specimen per plate. If
we try two or more specimens in a single 10 cm plate, this method is not suitable.
2. T-Streaking
It is another method of streaking where the agar Petri plate is divided into three sections and
each section is streaked. Hence, this method is also known as the “three-sector streak”
method.
The media is divided into three sections by drawing a letter “T” and each adjacent section is
streaked sequentially. By the time the final section is being streaked, the inoculum is diluted
to the point to give rise to isolated colonies following the incubation. Mostly discontinuous
fashion of streaking is followed; however, a continuous fashion can also be used in the very
dilute specimen.
3. Continuous Streaking
It is another commonly followed method where an inoculum is evenly distributed in a single
continuous movement from starting point to the center of the plate. There is no need to
divide the plate and sterilize the loop during the process. It is easy and quick; however, the
problem is that we can use it only if the inoculum is either very diluted or we just have to
propagate pure culture rather than isolate one.
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We can divide the 10 cm Petri plate into different sections (mostly 2 to 6), and in each section,
we can streak different specimens following this method. Hence, it is used in the clinical
laboratory to culture urine, sputum, pus, etc. if multiple samples have arrived at a single time.
This will allow us to save media and get maximum output using a minimum resource.
4. Radiant Streaking
It is another method of streaking where the inoculum is first streaked at one edge and spread
in vertical lines above the edge. Finally, the vertical lines are cross streaked diagonally. This
method is suitable to propagate pure culture, and also in the case of a dilute specimen.
5. Semi-quantitative Streaking
It is routinely followed in urine culture. It is a modified form of continuous streaking. In this
method, a calibrated loop (usually a loop of 1 or 2μl) is used to streak a certain volume of the
liquid specimen. A loopful of the specimen is streaked in a horizontal line in the middle of the
Petri plate, and the specimen is spread all over the plate in a single continuous back and forth
movement. This method allows us to approximately quantify the viable load (in a range, not
an exact number) as well as get the pure culture in a single go.
6. Zigzag Streaking
It is another form of continuous streaking where a loopful of the specimen is streaked all over
the plate in a zigzag pattern in a single continuous movement. It is commonly done to
propagate the pure culture and culture them in large quantities.
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This is a sterile tool used to streak the specimen over the surface of culture media. The tools
used for streaking are cotton swabs, inoculating loop (both metal and plastic), toothpicks, and
wooden or metal or plastic sticks/wires. The most commonly used one is inoculating loop
(nichrome wire loop).
2. Sample culture
Sample bacteria may be in the form of suspension, liquid broth, or colonies over solid media.
The sample is picked by using an inoculating loop and transferred over the surface of fresh
culture media to perform streaking.
Specific culture media is used for the isolation and differentiation of suspected (or specific)
bacteria. The culture medium is a solid agar medium that is pre-solidified before use.
A Bunsen burner is used to sterilize the loop and also to create a sterile zone around the
flame. Besides, other chemicals, sterilizing materials, and laboratory apparatus are also
required.
1. Arrange all the requirements, put on the PPE, sterilize the work surface, and allow all the
samples and media to come to room temperature if were refrigerated.
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2. If the sample is very concentrated then dilution can be helpful to get the isolated colonies.
(But it is not compulsory as the sample will be diluted during the streaking process.)
3. Sterilize the inoculating loop by flaming and allow it to cool. Pick a small portion of the
isolated colony. (if the sample is in the suspension then take a loopful of the sample)
The inoculating procedure is different according to the method of streaking, let us deal with
each type:
(if you are left-handed, hold the plate in your right hand)
2. The sample is spread over about 1/4th of the media in the Petri plate from the rim to the
center of the plate using a rapid, gentle, back and forth motion.
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(For ease, a beginner can draw two diameters intersecting each other diagonally at the back of
the petri dish to divide the media into 4 equal sections)
3. Re-flame the loop and allow it to cool. Turn the Petri plate by 90° anticlockwise, and place
the loop to the last streaks of the first quadrant. Move the loop back and forth to spread
the inoculum over the last half of the streaks in the first quadrant into the empty second
quadrant.
(Be sure not to move the loop to the streaks in the first half of the first quadrant.)
4. Repeat the process (iii) for streaking the third quadrant and the fourth quadrant.
5. For the fourth quadrant similar step can be followed. However, many people prefer to draw
a few (6 to 7 streaks) well-separated streaks by touching the second half of streaks in the
third quadrant. Also, some prefer to make the final streak in a zigzag fashion making a tail.
(In a discontinuous fashion, the loop is sterilized after streaking each quadrant. In a continuous
fashion, there is no need to flame the loop after streaking each quadrant. But, this is preferred
only if the sample is very dilute.)
T-Streaking Procedure
1. Lift the Petri plate in your left hand and hold it at an angle of 600. (if you are left-handed,
hold the plate in your right hand)
2. The sample is spread over about 1/3rd of the media in the Petri plate from the rim to the
center of the plate using a rapid, gentle, back and forth motion.
(For ease, a beginner can draw a letter “T” at the back of the petri dish to divide the media into
3 sections)
3. Re-flame the loop and allow it to cool. Turn the Petri plate by 900 anticlockwise, and place
the loop to the last streaks of the first quadrant. Move the loop back and forth to spread
the inoculum over the last half of the streaks in the first quadrant into the empty second
quadrant.
(Be sure not to move the loop to the streaks in the first half of the first quadrant.)
4. Repeat the process (iii) for streaking the third quadrant. As in quadrant streaking, you can
follow any one of the streaking patterns at the 3rd quadrant.
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1. Lift the Petri plate in your left hand and hold it at an angle of 600. (if you are left-handed,
hold the plate in your right hand)
2. Place the loop at one end of the plate and start streaking the inoculum from that point in a
continuous movement to the center of the plate.
3. Rotate the plate by 1800 and without sterilizing the loop, follow the step (ii) to streak the
remaining half of the plate.
5. Label at the edge of the bottom of the plate with the date, name, sample ID, and other
required information.
6. Incubate the plate in an inverted position under suitable incubation conditions (mostly for
24 hours at 370C).
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[Exception: in some cases where colony characters of two or more bacterial species are the
same, all the colonies may look alike even if they are of a different individual.]
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References
1. Sanders E. R. (2012). Aseptic laboratory techniques: plating methods. Journal of visualized
experiments : JoVE, (63), e3064. https://doi.org/10.3791/3064
2. Textbook of Microbiology and Immunology (2012), 2nd Editions. Subah Chandra Parija.
ISBN: 978-81-312-2810-4
3. Practical Handbook of Microbiology, 2nd Edition.Edited by Emanuel Goldman and Lorrence
H. Green. CRC Press. Taylor & Francis Group. 6000 Broken Sound Parkway NW, Suite 300.
6000 Broken Sound Parkway NW, Suite 300.
4. VAN Soestbergen, A. A., & Lee, C. H. (1969). Pour plates or streak plates?. Applied
microbiology, 18(6), 1092–1093. https://doi.org/10.1128/am.18.6.1092-1093.1969
5. Streak Plate Method – Explained – Laboratoryinfo.com
6. Streak Plate Method: Patterns, Procedure, Principle (microbiologynote.com)
7. Streak Plate Method: Principle, Procedure, Uses • Microbe Online
8. Microbiological Streaking Repair – iFixit
9. Streak Plate Method: Principle, Purpose, Procedure, And Results – BIOCHEMINSIDER
10. Streak Plate Method Principal and Types – RBR Life Science
11. Streak Plate Method (Procedure) : Microbiology Virtual Lab I : Biotechnology and
Biomedical Engineering : Amrita Vishwa Vidyapeetham Virtual Lab
12. Streak plates (wisc.edu)
13. Streak Plate – Virtual Interactive Bacteriology Laboratory (msu.edu)
14. Streaking Agar Plates: 4 Quadrant Streak Method – Microbiology learning: The “why”ology
of microbial testing (weebly.com)
15. Streak Plate Technique For Isolation of Microorganism | Culture Methods
(paramedicsworld.com)
16. What is the purpose of streak plate method? (askinglot.com)
17. Streaking method – Labster Theory
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Dianne Miller
July 21, 2022 at 11:26 AM
Thanks for explaining. Streak Plate method testing means a long, thin line, and the
streak plate method is a microbiological culture technique where a sample is spread
in a petri dish in the form of a long, thin line over the surface of solid media. I like that
you discuss the process streak plate method as a microbiological laboratory
technique of isolating pure cultures and getting well-isolated colonies of bacteria
from a mixed population. It is mainly used to obtain pure bacteria cultures; however,
this method can also isolate yeasts. It is one of microbiology’s most commonly used
aseptic techniques to isolate and propagate bacteria. It is a mechanical isolation
technique in microbiology, widely known as the streaking method.
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Chongo
March 31, 2020 at 7:33 AM
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Thanks a lot your website is very useful for me I was able to make my own notes and
easy to understand. Share pdf as well.
Reply
Gopal sharma
September 1, 2019 at 10:54 PM
Thanks
Reply
Vigneshwaran.K
July 13, 2019 at 4:55 PM
Reply
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