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The main thing that distinguishes the speech of different characters is what they say, not how they say it. If you understand the motives and the fears of every characters in your scene, and if you...
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#3: Attribution notice added
Source: https://writers.stackexchange.com/a/28337 License name: CC BY-SA 3.0 License URL: https://creativecommons.org/licenses/by-sa/3.0/
#2: Initial revision
The main thing that distinguishes the speech of different characters is what they say, not how they say it. If you understand the motives and the fears of every characters in your scene, and if you make sure that every word they speak proceeds from their fears and desires, then their speech will seem distinct and authentic. Only Fred would say that is fundamentally more distinguishing than only Fred would say it like that. Secondly, remember that fictional dialogue is not like regular speech. Regular speech relies on a bunch of clues and cues that are not present on the page. The words themselves therefore have to do more of the work. Some authors do attempt to record all of the clues and cues of real speech but it does not usually work well because the way we receive and interpret such clues and cues in everyday life is largely subconscious. An actor has access to the communication channel, an author of prose does not. Third, remember that regular speech is often disorganized and hard to follow because people don't always express themselves well or think through what they want to say. Reproducing all of this in fiction is usually tedious. The point of most dialogue is not to paint a portrait of human speech as it is in the wild but to advance the plot of the story, to reveal motivation or conflict, or to pass information. Reproducing the chaos and hesitancy of real speech would simply get in the way of the job you need the dialogue to do. Finally, in real life, most people who converse on a regular basis have very similar patterns of speech. We are social and tribal animals with a natural tendency to imitate each other as a sign or our willingness to fit in with the group. (You can observe this behavior very easily in children, especially around three to five, when imitative play and mimicry of speech and action is a huge part of their daily activities.) Where authors do reach for different speech patterns in fiction, therefore, is when the introduce a stranger into the mix. The difference in speech patterns is a way of emphasising their otherness. Thus fictional dialogue is an artifice that is often very different in sound and in purpose from ordinary human speech. For the most part, therefore, you distinguish characters more by what they say than by how they say it, you tidy up the untidiness of human diction, and you say more in words and less in actions than occurs in real human speech.