First Minister Nicola Sturgeon is set to clash with former London mayor Boris Johnson as part of a live television debate on STV.
The ITV Referendum Debate will feature Sturgeon, Angela Eagle and Amber Rudd for the Remain campaign.
Representing the arguments for leaving the European Union will be Boris Johnson, Andrea Leadsom, and Gisela Stuart, from Vote Leave.
They will appear on the event live between 8-10pm on Thursday. The two-hour debate will be moderated by ITV News's Julie Etchingham.
This will be the second of ITV's live European referendum programmes, and will be broadcast from London.
The politicians will each in turn make short opening statements at the beginning of the programme and closing statements at the conclusion of the debate.
During the two-hour encounter, they will address questions posed by the studio audience.
The format will allow each politician to give a short uninterrupted answer to each question, before the floor is opened up to free flowing debate.
The audience will be asked for their questions, as well as viewers, who have been asked to submit questions via firstname.lastname@example.org.
The ITV Referendum Debate will give viewers the chance to see senior politicians engaging directly in debate on the significant issues, ahead of Britain's critical vote to leave or to remain in the European Union on June 23.
On Tuesday, STV will broadcast a debate between the Prime Minister and UKIP leader Nigel Farage. Cameron and Farage Live: The EU Referendum, will air at 9pm on Tuesday, June 7.
Julie Etchingham will also present this hour-long live programme and both politicians will answer questions from a studio audience. Nigel Farage will take questions first, followed by David Cameron. The audience will pose their questions directly to the politicians.