
Amy C. answered 10/07/20
PhD in Musicology (Music History)
This is a great question. The answer is that they used their ears! They tuned notes in relation to each other, and they tuned instruments in relation to each other. They knew pitches (=frequencies, even if they didn't have that scientific knowledge) and which ones sounded good together to them, and they used that combination of notes.
Musicians still do this today. If you go to an orchestra concert or a band concert, right before the ensemble begins its program, you will hear all the instruments stop their warmup, and an oboe will play a note (usually an A). The musicians will start by matching their A to that of the oboe, and then they will test out other notes in relation to their freshly tuned A--this is especially important for string instruments where they have to tune more than one string. Similarly, piano tuners used to use tuning forks (and perhaps they still do) to get the pitch of one string down, and then they have to tune all the others in relation to that.
What the musicians are listening for is purity of sound. When you combine two frequencies that are just shy of matching, the sound waves combine in a way that makes the sound quality harsh, and even produces variations in volume called "beats." Two instruments are in tune with each other when they can play the same note without any resulting beats.
And the process is similar for testing out other notes in relation to a tuned note. This depends in part on the harmonic series, the fact that any note produced also produces other notes. So, for example, if you play a low C on the piano, it also produces a softer C an octave above that, an even softer G above that, the next C, the next E, and it continues on. Those extra pitches are absorbed into the sound, and they all combine to produce that low C sound. You will note that C-G, the interval of a fifth, produces the second harmonic, which is more easily heard than the harmonics after that. This is why most tuning in Western music is done in fifths--violins have strings tuned to G, D, A, and E. If a fifth is out of tune, it will produce a similar effect to if a unison note is out of tune--harsh quality, beats. Guitar tuning is based mostly on fourths, which is the third harmonic.
And in fact, a lot of musicians prefer to tune by ear than by using a tuner, since if there are multiple musicians using multiple tuners, the pitch might not match exactly. Human/mechanical error and all that.
Please let me know if you want to know more!