Mauricio Pochettino was optimistic Kane would be fit to play some part in Wednesday's tie at Wembley.
And the signs looked good in training as the smiling striker was involved in drills after an indoor gym session. Erik Lamela also took part.
Harry Kane was back in action for Tottenham as they prepared to take on Real Madrid
Kane gestures to reassure Tottenham team-mates as he works ahead of the Real tie
The 24-year-old appeared to tell one Tottenham player 'I'm alright' during the session and gave an OK gesture to reassure the team-mate.
Kane was forced to sit out Tottenham's last two matches after picking up a hamstring problem in their 4-1 Premier League win over Liverpool.
Pochettino provided an update on Kane's fitness at his press conference at the Tottenham training ground.
'We're very confident,' Pochettino said when asked about Kane's availability. 'He's on the list. Maybe it's possible for him to play but I don't want to say 100 per cent'.
'He trained but it is important we make the right decision,' Pochettino added. 'With the doctors and the player, we will take the best decision for everyone. We must analyse and take the best decision for everyone.
Kane looked in good spirits as he got involved with drills following an indoor gym session
Kane has been unavailable for Tottenham's past two matches with a hamstring problem
Mauricio Pochettino said he would analyse Kane's fitness with Tottenham's club doctors
'We will assess him and then we will make the best decision. My feeling is good. But it's not my feeling, it's Harry Kane's feeling.'
Kane trained alongside Lamela, who hasn't featured for Tottenham since October 2016 after suffering repeated hip problems.
Key men Christian Eriksen, Dele Alli and Son Heung-min were also present during the session as Tottenham bid to maintain their strong start to the Champions League campaign.
Alli was suspended for Tottenham's 1-1 draw with Madrid at the Bernabeu earlier this month but is available once again for selection ahead of the Wembley clash.
Erik Lamela hasn't played since October 2016 after suffering a string of hip problems
Kane leads out his Tottenham team-mates for Tuesday afternoon's open training session
Christian Eriksen and Eric Dier were in action as Tottenham prepared to take on the holders
Kane was all smiles as he bids to make his first appearance since the 4-1 thrashing of Liverpool
Tottenham took the lead after Kane's flick took a deflection of Raphael Varane resulting in an own goal but Madrid equalised through Cristiano Ronaldo's penalty.
The away draw at Madrid followed two consecutive victories to start Tottenham's European campaign, against Borussia Dortmund and APOEL Nicosia.
Tottenham have lost both their matches without Kane since the striker suffered the hamstring problem towards the end of the win over Liverpool.
They threw away a two-goal lead to lose 3-2 to West Ham in the Carabao Cup before a 1-0 Premier League loss at Manchester United on Saturday.
Dele Alli (left) and Juan Foyth (centre) take part in a drill on Tuesday
Kane shared a joke with Son Heung-min as Tottenham geared up for Wednesday's match
Kyle Walker-Peters hasn't featured since the 4-0 win over Huddersfield on September 30