"France 24 has officially filed a complaint to the Paris public prosecutor regarding identity theft and impersonation," said a statement from the channel.
The channel explained that it had invited ambassador Lamia Shakkur to participate in a live debate programme after first contacting the embassy by phone and email.
France 24 declared that the embassy provided a telephone number and photograph which were presented as being those of the ambassador and which were then used live on-air.
In Tuesday's live interview the woman introduced as ambassador Lamia Shakkur surprised viewers by announcing on air that she was resigning in protest at "the cycle of violence" alluding to the actions of Syrian authorities against protestors.
Reuters news agency subsequently issued dispatches citing the Syrian embassy and confirming the resignation.
Lamia Shakkur then went on other television channels, denouncing what she said was an attempt to impersonate her, though she refused the opportunity to appear on France 24 to explain what she thought had happened.
"France 24 has no option but to take Ms Shakkur's denials seriously," the channel said in a press release on Thursday, adding that it had "commissioned comparative analyses which show that the voice heard during the interview conducted on France 24 on Tuesday evening is different from the voice which later issued the denial broadcast on BFM TV," another channel which Shakkur spoke to.
The statement declared that both France 24 and ambassador Lamia Shakkur were victims in the affair, and that the channel expected that she and the embassy would give their total support to the legal action, as well as their complete cooperation in investigations into the case.