Investors seemed concerned that the average number of monthly users had climed just 30% to 241 million compared to the same quarter the previous year.
This saw the firm's share price slide more than 17% to $54.24 (€40.11).
It came despite results showing revenues in the quarter ending December 31 doubled to a better-than-expected $242.6m (€179.41m).
In its annual earnings report it showed its total revenue for last year was $665m (€492m), more than double the $316m (€234m) of 2012.
However, chief executive Dick Costolo said the site would only get better.
"Twitter finished a great year with our strongest financial quarter to date," he said.
"We are the only platform that is public, real-time, conversational and widely distributed and I'm excited by the number of initiatives we have under way to further build upon the Twitter experience."
Twitter was launched seven years ago.