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Direct Counting of Cells, Spores,Viable Plate Count

Direct Counting of Cells, Spores,Viable Plate Count

There are several methods that can be used for counting the cells and spores of bacteria and other microbes. These include direct count, plate count, and most probable number (MPN) determinations.

(i) Direct count

The density of cells, spores etc. of microorganisms can be found out by counting their number in a unit volume. The simplest technique is to use a special counting chamber, of a type that used in a haemocytometer for counting the blood cells. The counting chamber is simply a ruled slide with a supported glass cover that holds a definite volume of fluid.

(ii) Viable plate count

This is one of the most common methods, for enumeration of bacteria. Serial dilutions of suspension of bacteria are plated onto a suitable solid medium. The method of serial dilution of sample has already been described earlier, where we could have 10-1 to 10-8 or more dilutions of the sample. Dilution procedure influences overall counting process. The suspension is either spread over the surface of growth medium or mixed with the agar prior to its solidification and then poured into the plate. The plates are incubated so that colonies ate formed. Multiplication of a bacterium on solid media results in the formation of a macroscopic colony visible to naked eye. It is assumed that each colony arises from an individual viable cell.

Total number of colonies are counted and this number multiplied by dilution factor to find out concentration of cells in the original sample. Counting plates should have 30-300 colonies at least. A major limitation in this method is selectivity.

The nature of the growth medium and the incubation conditions determine which bacteria can grow and thus be counted. Viable counting measures only those cells that are capable of growth on the given medium under the set of conditions used for incubation. Sometimes cells are viable but non culturable

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