Microbiology 23 = Isolation of Pure Culture (Part- 02) | Pour Plate method | Spread Plate Method

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Pour plate method is usually the method of choice for counting the number of colony-forming bacteria present in a liquid specimen. In this method, a fixed amount of inoculum (generally 1 ml) from a broth/sample is placed in the centre of sterile Petri dish using a sterile pipette. Molten cooled agar (approx. 15mL) is then poured into the Petri dish containing the inoculum and mixed well. After the solidification of the agar, the plate is inverted and incubated at 37°C for 24-48 hours.
Microorganisms will grow both on the surface and within the medium. Colonies that grow within the medium generally are small in size and maybe confluent; the few that grow on the agar surface are of the same size and appearance as those on a streak plate. Each (both large and small) colony is carefully counted (using magnifying colony counter if needed). Each colony represents a “colony-forming unit” (CFU).
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. 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.
Use -
(1) It is used for viable plate counts, in which the total number of colony forming units on a single plate is enumerated.
(2) It is used to calculate the concentration of cells in the tube from which the sample was plated.
(3) Spread plating is routinely used in enrichment, selection, and screening experiments.
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