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The count represents the number of colony forming units (cfu) per g (or per ml) of the sample. A TVC is achieved by plating serial tenfold dilutions of the sample until between 30 and 300 colonies can be counted on a single plate. The reported count is the number of colonies counted multiplied by the dilution used for the counted plate
The pour plate technique is the typical technique used to prepare plate count agars. Here, the inoculum is added to the molten agar before pouring the plate. The molten agar is cooled to about 45 degrees Celsius and is poured using a sterile method into a petri dish containing a specific diluted sample.
The membrane filter is then placed onto Soybean-Casein Digest Agar and incubated in order to be able to determine the total aerobic microbial count (TAMC). [4] In the Plate Count Method, the sample of drug product to be tested and Soybean-Casein Digest Broth is poured into a Petri dish. [4] The Petri dish is then incubated.
Colony-forming units are used to quantify results in many microbiological plating and counting methods, including: The pour plate method wherein the sample is suspended in a Petri dish using molten agar cooled to approximately 40–45 °C (just above the point of solidification to minimize heat-induced cell death).
The plate count method relies on bacteria growing a colony on a nutrient medium so that the colony becomes visible to the naked eye and the number of colonies on a plate can be counted. To be effective, the dilution of the original sample must be arranged so that on average between 30 and 300 colonies of the target bacterium are grown.
A third technique is using sterile glass beads to plate out cells. In this technique, cells are grown in a liquid culture, in which a small volume is pipetted on the agar plate and then spread out with the beads. Replica plating is another technique used to plate out cells on agar plates. These four techniques are the most common, but others ...
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Determining the viable cell count is important for calculating dilutions required for the passaging of cells, as well as determining the size and number of flasks needed during growth time. It is also vital when seeding plates for assays, such as the plaque assay , [ 2 ] because the plates need a known number of live replicating cells for the ...