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Newton's main interest at the time of his appointment was optics, and for several years the lectures required of him by the professorship were devoted to this subject. In a letter of 1672 to the secretary of the Royal Society, he says that in 1666 he had bought a prism "to try therewith the celebrated phenomena of colours." He continues, "In order thereto having darkened the room and made a small hole in my window-shuts to let in a convenient quantity of the Suns light, I placed my prism at its entrance, that it might be thereby refracted to the opposite wall." He had been surprised to see the various colors appear on the wall in an oblong arrangement (the vertical being the greater dimension), "which according to the received laws of refraction should have been circular." Proceeding from this experiment through several stages to the "crucial" one, in which he had isolated a single ray and found it unchanging in color and refrangibility, he had drawn the revolutionary conclusion that "Light itself is a heterogeneous mixture of differently refrangible rays."

These experiments had grown out of Newton's interest in improving the effectiveness of telescopes, and his discoveries about the nature and composition of light had led him to believe that greater accuracy could not be achieved in instruments based on the refractive principle. He had turned, consequently, to suggestions for a reflecting telescope made by earlier investigators but never tested in an actual instrument. Being manually dexterous, he built several models in which the image was viewed in a concave mirror through an eyepiece in the side of the tube. In 1672 he sent one of these to the Royal Society.

Newton felt honored when the members were favorably impressed by the efficiency of his small reflecting telescope and when on the basis of it they elected him to their membership. But when this warm reception induced him to send the society a paper describing his experiments on light and his conclusions drawn from them, the results were almost disastrous for him and for posterity. The paper was published in the society's Philosophical Transactions, and the reactions of English and Continental scientists, led by Robert Hooke and Christiaan Huygens, ranged from skepticism to bitter opposition to conclusions which seemed to invalidate the prevalent wave theory of light.

At first Newton patiently answered objections with further explanations, but when these produced only more negative responses, he finally became irritated and vowed he would never publish again, even threatening to give up scientific investigation altogether. Several years later, and only through the tireless efforts of the astronomer Edmund Halley, Newton was persuaded to put together the results of his work on the laws of motion, which became the great Principia.