In 1876, former California governor Leland Stanford bought the Palo Alto Farm as his racehorse breeding ground. Stanford and Charles Marvin, the Superintendent of Palo Alto Farm, created new methods for training horses, which Marvin described in his book 'Training the Trotting Horse' (1890) - which includes a picture of the famous trotter 'Palo Alto' after who the farm (and city) were named. The horse had in turn been named after the first battlefield victory in the Mexican War (1846). The farm was the site for Stanford University which was founded in 1885 as a memorial to Stanford's son who died of typhoid.

Stanford is reputed to have made a bet with and business associates James R. Keene and Frederick MacCrellish, that when a racehorse was at full gallop there were times when all four legs were off the ground. The stake was high, perhaps $25,000, a huge fortune at the time, and there seems to be no firm evidence that it was ever made or paid. This was a time when the horse was still vital both to the economy as a means of power and transport as well as for activities such as racing, and when scientific studies were beginning to improve the breeding and training of horses. A study such as this of the horse in motion was very much the kind of thing a wealthy horse breeder might decide worth undertaking.

To win the bet Stanford persuaded Muybridge to make a photographic study of motion of horses. Given that the normal exposures with the wet plate process, then the fastest known materials, were several seconds, this was a daunting task. Muybridge told Leland it was beyond what photography could currently do, but Leland replied that if Muybridge put his mind to it, he would be able to do it, and eventually he was proved correct, but it was a long, expensive and difficult job, probably costing twice the stake to win the bet for Stanford, and taking over 10 years.

Since he knew it was impossible to take normal photographs with a sufficiently rapid shutter speed, Muybridge tried to record silhouettes without any shadow detail. He photographed the horse against a white wall lit by strong sunlight, using relatively wide lens apertures. The plates were still greatly underexposed, but extended development produced faint silhouettes which could be printed and retouched.

The horse was set to gallop past a series of cameras facing the white background. Each camera had a specially designed rapid shutter connected to a string across the track; as the horse galloped past it broke the string and released the shutter, thus taking its own picture. Dark lines on the background enabled the height of the hooves above the ground to be easily determined.

Many refused to believe they were true. Muybridge was able to show that they were by viewing or projecting the pictures in rapid succession, when the horse's motion could be reproduced. He was thus one of the pioneers of the movies . In the 1890s Edison bought the rights to animate and use a hundred of his plates.

Muybridge was able to extend his work at the University of Pennsylvania from 1884-1887. The newly introduced dry plates, first mass produced around 1880, were much faster than wet plates, making his work simpler and producing more detail. Banks of cameras, now fitted with clockwork controlled shutters, were used to photography human and animal movements from several viewpoints.