A novel procedure for CFU plating and counting

Abstract

We present a novel method to automate plating for the Colony Forming Unit (CFU) counting procedure. The apparatus we developed for the application of this method is based on motorized stages and a syringe in order to spread fine drops of a liquid containing a solution of interest on a plate without direct contact with the surface. The apparatus can be used in two different modes. In the the first method which follows the same principle as the classical CFU count, fine drops of liquid are deposited homogeneously on an agar plate and microorganisms are allowed to form colonies. In the second, novel method that we call P0, isolated drops of the order of 10 μL containing both the microbes and the nutrient medium are deposited directly on a regular grid on a hard surface (plastic or glass); after incubation, drops that show no sign of growth inside are used to determine the microbes concentration. This new method removes the need for preparing agar surfaces and allows for easy disposal of waste and reuse of consumables. The apparatus is simple to build and use, the plating is fast and the CFU count in both type of plating is extremely reproducible and robust.