The air-pump was invented shortly before the foundation of the Royal Society, and its members made many experiments with this new means of isolating an agent and thus discovering its potentialities. For example, live animals were put into the receiver, and the air exhausted, with the result that they quickly died. The absence of the air being the sole difference, it was thus proved to be indispensable to life. But air is a composite agent, and when means were contrived of separating its components, the effects of oxygen alone and of carbonic acid alone were experimentally determined. A good example of the difficulty of excluding agencies other than those we are observing, of making sure that none such intrude, is found in the experiments that have been made in connexion with spontaneous generation. The question to be decided is whether life ever comes into existence without the antecedent presence of living germs. And the method of determining this is to exclude all germs rigorously from a compound of inorganic matter, and observe whether life ever appears. If we could make sure in any one case that no germs were antecedently present, we should have proved that in that case at least life was spontaneously generated. The difficulty here arises from the subtlety of the agent under observation. The notion that maggots are spontaneously generated in putrid meat, was comparatively easy to explode. It was found that when flies were excluded by fine wire-gauze, the maggots did not appear. But in the case of microscopic organisms proof is not so easy. The germs are invisible, and it is difficult to make certain of their exclusion. A French experimenter, Pouchet, thought he had obtained indubitable cases of spontaneous generation. He took infusions of vegetable matter, boiled them to a pitch sufficient to destroy all germs of life, and hermetically sealed up the liquid in glass flasks. After an interval, micro-organisms appeared. Doubts as to the conclusion that they had been spontaneously generated turned upon two questions: whether all germs in the liquid had been destroyed by the preliminary boiling, and whether germs could have found access in the course of the interval before life appeared. At a certain stage in Pouchet's process he had occasion to dip the mouths of the flasks in mercury. It occurred to Pasteur in repeating the experiments that germs might have found their way in from the atmospheric dust on the surface of this mercury. That this was so was rendered probable by his finding that when he carefully cleansed the surface of the mercury no life appeared afterwards in his flasks.