- Advantage And Disadvantage Of The Serial Dilution Agar Plate Procedure Form
- Advantages Of Serial Dilution-agar Plate Procedure
- Advantage And Disadvantage Of The Serial Dilution Agar Plate Procedure Instructions
Medium: Plate count agar or Nutrient agar; Procedure for Spread Plate Technique: A: Serial Dilution. Prepare a series of at least 6 test tubes containing 9 ml of sterile distilled water. Using a sterile pipette,add 1ml of sample in the first tube of the set.Label it as 10-1; Mix the contents well by swirling the tube upside down few times. Serial dilutions & 'spread plate technique'? If you were to directly take a given serial dilution and do a count under a microscope what would be the advantages and disadvantages of this method versus carrying out the serial dilution-agar plate procedure to count the number of 'cells'?
Spread plate technique is the method of isolation and enumeration of microorganisms in a mixed culture and distributing it evenly. The technique makes it easier to quantify bacteria in a solution.
Principle of Spread Plate Technique
The spread plate technique involves using a sterilized spreader with a smooth surface made of metal or glass to apply a small amount of bacteria suspended in a solution over a plate. The plate needs to be dry and at room temperature so that the agar can absorb the bacteria more readily. A successful spread plate will have a countable number of isolated bacterial colonies evenly distributed on the plate.
Procedure of Spread Plate Technique
- Make a dilution series from a sample.
- Pipette out 0.1 ml from the appropriate desired dilution series onto the center of the surface of an agar plate.
- Dip the L-shaped glass spreader into alcohol.
- Flame the glass spreader (hockey stick) over a Bunsen burner.
- Spread the sample evenly over the surface of agar using the sterile glass spreader, carefully rotating the Petridish underneath at the same time.
- Incubate the plate at 37째C for 24 hours.
- Calculate the CFU value of the sample. Once you count the colonies, multiply by the appropriate dilution factor to determine the number of CFU/mL in the original sample.
Uses of Spread Plate Technique
- It is used for viable plate counts, in which the total number of colony forming units on a single plate is enumerated.
- It is used to calculate the concentration of cells in the tube from which the sample was plated.
- Spread plating is routinely used in enrichment, selection, and screening experiments.
Limitations of Spread Plate Technique
- Strick aerobes are favored while microaerophilic tends to glow slower.
- Crowding of the colonies makes the enumeration difficult.
References
- M. J. Pelczar
- Mackie and McCartney. Practical Medical Microbiology
- Bailey & Scott’s Diagnostic Microbiology.
- Practical Microbiology. Pradeep Kumar Sharma.
Agar dilution is one of two methods (along with Broth Dilution) used by researchers to determine the Minimum Inhibitory Concentration (MIC) of antibiotics. It is the dilution method most frequently used to test the effectiveness of new antibiotics when a few antibiotics are tested against a large panel of different bacteria.[1]:149[2]
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![Serial Serial](https://www.coursehero.com/thumb/f8/5e/f85e0b13fac17238375a02ed5718c6b5eaa112f7_180.jpg)
Process[edit]
The antibiotic to be tested is diluted with water to produce a series of concentrations. An appropriate volume is then combined with melted agar to produce plates in which the final antibiotic concentrations represent a 2-fold dilution series. After this, bacteria prepared to a standard concentration are added as a spot to each plate, with 104 colony forming units (CFU) per spot. This technique allows for replicate spots of one bacterial type to be tested [3] or spots of different bacteria so that the MIC of the antibiotic against multiple types of bacteria can be tested. Necessary controls include a control plate that does not receive any antibiotics and bacterial spread plates demonstrating that the bacterial inocula are in the correct concentration range. The dilution plates are then incubated at a temperature of 37 degrees Celsius.[4] The plates are then incubated for sixteen to eighteen hours, although incubation time may be less for bacteria populations that divide quickly.[1]:374 Elfbot 8.60 crack tibia laner download. After incubation, the plates are examined to determine if bacterial growth has occurred in the inoculated spots. The lowest concentration of antibiotics that prevented bacterial growth is considered to be the minimum inhibitory concentration of that antibiotic against that bacterium.[4]
Advantage And Disadvantage Of The Serial Dilution Agar Plate Procedure Form
Advantages[edit]
![Plate Plate](https://i1.rgstatic.net/publication/225056141_Aseptic_Laboratory_Techniques_Plating_Methods/links/562155a308ae93a5c927ded4/largepreview.png)
Agar dilution is considered to be the gold standard of susceptibility testing, or the most accurate way to measure the resistance of bacteria to antibiotics. The results of agar dilution are easily reproduced and they can be monitored at a much cheaper cost than what is required of other dilution methods. Additionally, up to thirty pathogen samples (plus two controls) can be tested at once, so agar dilution is useful for batch tests.[1]:149
Disadvantages[edit]
Advantages Of Serial Dilution-agar Plate Procedure
Each dilution plate in agar testing has to be manually inoculated with the pathogen to be tested, so agar dilution testing is both labor-intensive and expensive.[1]:149 Unlike broth microdilution tests, agar dilution cannot be used to test more than one antibiotic at a time.[5]
References[edit]
- ^ abcdLorian, Victor (2005). Antibiotics in Laboratory Medicine. Lippincott Williams & Wilkins.
- ^Wiegand, Irith; Hilpert, Kai; Hancock, Robert E. W. (17 January 2008). 'Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances'. Nature Protocols. 3: 163–75. doi:10.1038/nprot.2007.521. PMID18274517.
- ^European Committee for Antimicrobial Susceptibility Testing of the European Society of Clinical Microbiology and Infectious Disease (September 2000). 'Determination of minimum inhibitory concentrations (MICs) of antibacterial agents by agar dilution'. Clinical Microbiology and Infection. 6 (9): 509–515. doi:10.1046/j.1469-0691.2000.00142.x.
- ^ abParija (2009). Textbook of Microbiology & Immunology. Elsevier India. p. 73.
- ^Lee, Mary (2013). Basic Skills in Interpreting Laboratory Data (5 ed.). ASHP. p. 723. Retrieved 16 November 2014.
Advantage And Disadvantage Of The Serial Dilution Agar Plate Procedure Instructions
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