Pasteur's experiments to prove these facts were extremely simple. He took a series of bulbs of about a quarter of a litre in capacity, and, after having half filled the bulbs with a putrescible liquid, he drew out the necks by means of the blowpipe, then he caused the liquid to boil for some minutes, and during the ebullition, while the steam issued from the tapering ends of the bulbs, he sealed them with the lamp. Thus prepared, the bulbs can be easily transported. As they are empty of air—that which they originally contained having been driven out with the steam—when the sealed end of a bulb is broken off, the air rushes into the tube, carrying with it all the germs which this air holds in suspension. If it is closed again immediately afterwards by a flame, and if the vessels are then left to themselves, it is easy to recognise those in which a change occurs. Now, Pasteur established that, in whatever place the operation might be carried on, a certain number of bulbs would escape alteration. They must not, however, be opened in a room after dusting the furniture or sweeping the floor, for in this case all the bulbs would become altered because of the great quantity of germs raised by the dusting and remaining suspended in the air.

Pasteur started for Arbois with a series of bulbs. Some he opened in the country far from all habitations; others he opened at the foot of the mountains which form the first range of the Jura; a series of twenty-four bulbs was opened upon Mount Poupet, at 850 meters above the level of the sea; and, lastly, twenty others were transported to the Montanvert, near the Mer de Glace, at an elevation of 2,000 meters. He afterwards brought his whole collection back to Paris, and in the month of November, 1860, deposited them on the table at the Academy of Sciences.

Of the twenty bulbs first opened in the country, eight contained organised productions. Of the twenty opened on the heights of the Jura, five only were altered, and of the twenty opened upon the Montanvert during a strong wind which blew from the glacier, one alone was altered.

If a similar series of experiments were made in a balloon, it would be found that the air of the higher atmosphere is absolutely free from germs. Care would, however, be necessary to prevent the introduction of dust particles, which the rigging and the aëronauts themselves might carry with them.

But we have not yet related all. So far, all these conclusive experiments had been made only on organic liquids, very putrescible it is true, but which had all been subjected to boiling or even to temperatures higher than 100 degrees Centigrade. The partisans of spontaneous generation might then be justified in saying that if the precaution had been taken of putting into contact with pure air natural organic liquids in a state compatible with the operations of animal and vegetable life, the results would have been different. Under such conditions, life would have appeared spontaneously in the production of microscopic organisms. None of Pasteur's opponents had formulated this argument; but Pasteur himself, who had within him an adversary always present, always on the alert, prepared to yield only to accumulated proofs, saw this objection. He was not satisfied until he had succeeded in completely refuting it. Having by means of ingenious experimental arrangements deprived some air of all living germs, he placed in contact with this pure air the most putrescible liquids, particularly venous blood, arterial blood, and urine. He took these liquids directly from the veins, the arteries, and the bladders of animals in full health. No alteration was produced. In due time a chemical absorption of small quantities of oxygen took place, but neither fermentation nor putrefaction, nor the smallest development of bacteria, of vibrios, or of mould. After this, Pasteur was able legitimately to exclaim in his celebrated lecture at the Sorbonne:

'There is not one circumstance known at the present day which justifies the assertion that microscopic organisms come into the world without germs or without parents like themselves. Those who maintain the contrary have been the dupes of illusions and of ill-conducted experiments, tainted with errors which they know not how either to perceive or to avoid. Spontaneous generation is a chimera.'

Pasteur was not alone in affirming this fixed conviction. With the authority of a judge delivering sentence in court, M. Flourens, permanent Secretary of the Academy of Sciences, pronounced these words before the whole Academy:

'As long as my opinion was not formed I had nothing to say; now it is formed and I can speak. The experiments are decisive. If spontaneous generation be a fact, what is necessary for the production of animalculæ? Air and putrescible liquids. Now Pasteur puts together air and putrescible liquids and nothing is produced. Spontaneous generation, then, has no existence. Those who still doubt have failed to grasp the question.'

But some adversaries remained incredulous. When Pasteur had announced the result of his experiments, and brought before the Academy his series of bulbs, Pouchet and Joly declared that if Pasteur had opened his bulbs in the Jura and on the Mer de Glace, they, on their part, had been on the top of the Maladetta, and had proved there the inexactitude of Pasteur's results.