By William Hewitt
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Extra info for Microbiological Assay. An Introduction to quantitative principles and Evaluation
The addition of 64 test solutions one by one to a plate results in a considerable time difference between addition of the first and the last. This represents a difference in diffusion time and leads to bias in resultant zone sizes. Brownlee et al describe a routine for application of test solutions designed to compensate for differences in diffusion time. Lees and Tootill (1955a-c) describe developments in the use of large plates and give examples of a variety of designs for different purposes.
According to Lees and Tootill (1955a) there is a further positional influence on zone size in large plates. This is a difference between inner zones and those in the rows and columns at the edge of the plate. Each inner zone is surrounded on all sides by reservoirs to which standard or test solutions have been applied. Water diffusing from these points keeps the moisture content in the center of the plate relatively higher than at the edge, where there is a tendency for the agar to dry out. Thus at the edges if the moisture content is lower, diffusion is faster and zones tend to be larger.
In such a design, each of the 16 n u m b e r s occurs once a n d only once in four of the eight rows a n d four of the eight columns. There are two sets of rows and t w o sets of columns, which are distinguished here by the letters A and B. In A rows, the n u m b e r s are 1, 3, 5, 7, 10, 12, 14, and 16, in Β rows, the n u m b e r s are 2, 4, 6, 8, 9, 11, 13, and 15, in A columns, the n u m b e r s are 1, 4, 5, 8, 10, 11, 14, a n d 15, and in Β columns, the n u m b e r s are 2, 3, 6, 7, 9, 12, 13, a n d 16.