Thus supported, Pouchet multiplied objections to the views of Pasteur, who had to meet every argument. Pasteur intended to narrow more and more the sphere of discussion. It was an ingenious operation to take the dusts from a cotton-wool filter, to disseminate them in a liquid, and thus to determine the alteration of that liquid; but the cotton wool itself was an organic substance and might be suspected. He therefore substituted for the cotton wool a plug of asbestos fibre, a mineral substance. He invented little glass flasks with a long curved neck; he filled them with an alterable liquid, which he deprived of germs by ebullition; the flask was in communication with the outer air through its curved tube, but the atmospheric germs were deposited in the curve of the neck without reaching the liquid; in order that alteration should take place, the vessel had to be inclined until the point where the liquid reached the dusts in the neck.
But Pouchet said, “How could germs contained in the air be numerous enough to develop in every organic infusion? Such a crowd of them would produce a thick mist as dense as iron.” Of all the difficulties this last seemed to Pasteur the hardest to solve. Could it not be that the dissemination of germs was more or less thick according to places? “Then,” cried the heterogenists, “there would be sterile zones and fecund zones, a most convenient hypothesis, indeed!” Pasteur let them laugh whilst he was preparing a series of flasks reserved for divers experiments. If spontaneous generation existed, it should invariably occur in vessels filled with the same alterable liquid. “Yet it is ever possible,” affirmed Pasteur, “to take up in certain places a notable though limited volume of ordinary air, having been submitted to no physical or chemical change, and still absolutely incapable of producing any alteration in an eminently putrescible liquor.” He was ready to prove that nothing was easier than to increase or to reduce the number either of the vessels where productions should appear or of the vessels where those productions should be lacking. After introducing into a series of flasks of a capacity of 250 cubic centimetres a very easily corrupted liquid, such as yeast water, he submitted each flask to ebullition. The neck of those vessels was ended off in a vertical point. Whilst the liquid was still boiling, he closed, with an enameller’s lamp, the pointed opening through which the steam had rushed out, taking with it all the air contained in the vessel. Those flasks were indeed calculated to satisfy both partisans or adversaries of spontaneous generation. If the extremity of the neck of one of these vessels was suddenly broken, all the ambient air rushed into the flask, bringing in all the suspended dusts; the bulb was closed again at once with the assistance of a jet of flame. Pasteur could then carry it away and place it in a temperature of 25-30° C., quite suitable for the development of germs and mucors.