But the position of Palavitalism changed in the second third of the nineteenth century. In 1833 appeared Johannes Müller's classical _Manual of Human Physiology_, in which the great biologist not only made a comparative study of the vital phenomena in man and the animals, but sought to provide a sound basis for it in all its sections by his own observations and experiments. It is true that Müller retained to the last (1858) the current idea of a vital force, as the supreme regulator of all the vital activities. However, he did not regard it as a metaphysical principle (like Haller, Kant, and their followers), but as a natural force, subject, like all others, to fixed chemical and physical laws, and subordinate to the whole. In his comprehensive study of every single vital function--the organs of sense and the nervous system, metabolism and the action of the heart, speech and reproduction--Müller endeavored above all to establish, by close observation of the facts and careful experiments, the regularity of the phenomena, and to explain their development by a comparison of the higher and lower forms. Hence Johannes Müller is wrongly described--as he has been of late--as a vitalist; he was rather the first physiologist to provide a physical foundation for the current metaphysical vitalism. He really gives an indirect proof of the reverse theory, as E. Dubois-Reymond rightly observed in his brilliant memorial speech. In the same way Schleiden (1843) cut the ground from under vitalism in botany. By his cell-theory (1838) he showed the unity of the multicellular organism to be the resultant of the functions of all the cells which compose it. The physical explanation of the vital processes and the rejection of Palavitalism were general in the last third of the nineteenth century. This was due most of all to the great advance in experimental physiology, which Carl Ludwig and Felix Bernard led as regards the animal body, and Julius Sachs and Wilhelm Preyer for the plant. While these and other physiologists used the remarkable results of modern physics and chemistry in the experimental study of the vital functions, and sought to determine their complicated course in terms of mass and weight and formulate their discoveries as mathematically as possible, they brought a great number of the wonders of life under the same fixed laws that were recognized in the physics and chemistry of the inorganic world. On the other hand, vitalism met with a powerful opponent in Charles Darwin, who solved, by his theory of selection, one of the most obscure biological problems, the constantly repeated question: How can we give a mechanical explanation of the orderly structures of the living being? How was this ingenious machine of the animal or plant body unconsciously produced by natural means, without supposing that some intelligent artificer or creator had deliberately designed and produced it?