Kezia Dugdale has moved to clarify her stance on independence after she said in an interview ''it's not inconceivable'' that she could support a future Yes vote if the UK leaves the EU.
Speaking to the Fabian Review, the Scottish Labour leader said she "would very much like'' to remain part of both the UK and EU.
Ms Dugdale was asked where her ''loyalty'' would be if there was an overall vote to leave in the EU referendum but the majority of Scots wanted to remain.
She told the Fabian Review: ''I've never contemplated that. I really wouldn't like to choose, because what I want to do is the best possible thing for Scotland. (I would be) putting Scotland first.''
When pushed on the topic and asked if she would ''argue, for Scotland's sake, against the UK Union", Ms Dugdale said: ''Possibly. It's not inconceivable.''
Ms Dugdale later moved to clarify her position in a statement, saying: "As I made clear in the leaders' TV debate this week, Labour has ruled out a second independence referendum. We won't introduce one in government and we would vote against one if it's introduced by any other party.
"I campaigned as hard as anybody to ensure that Scotland remained part of the UK. The collapse in the oil price showed that the best way to secure our public services is to stay in the UK.
"I would vote to stay in the UK in any future referendum."
Independence re-emerged as an election issues during a debate this week.
During the STV programme, Nicola Sturgeon said any future referendum decision was "in the hands of the people", while Tory leader Ruth Davidson accused the First Minister of disrespecting the 2014 poll.
Ms Davidson said Scottish Labour ''simply cannot be trusted to defend the decision of two million Scots to stay part of the UK''.
''The idea that Scotland's place in the United Kingdom is in some way dependent on Britain's membership of the EU is offensive," the Tory leader added.
"Scotland helped build the UK and is an integral part of it - confirmed by the referendum vote just 18 months ago.''
Scottish Liberal Democrat leader Willie Rennie said: "Only the Scottish Liberal Democrats are now unambiguously in favour of Scotland's place in the United Kingdom and the European Union.
"With Labour confusion over independence and the Conservative division over Europe, the SNP are rubbing their hands with glee at the prospect of another independence debate."