Blues boss believes his side must be at the top of its game if it is to overcome the current Champions League holders and is confident of handling Messi at Stamford Bridge.
The manager hailed the spirit in the Blues' dressing room following their 5-1 FA Cup semifinal win over Tottenham on Sunday, and the Italian believes the Catalans do not like playing at Stamford Bridge and that his side can prove to be awkward opposition for the Spanish giants.
"Certainly, the spirit in the group is very high and very good. I don't think we could have had a better game on Sunday coming into this one. We have a good history against Barcelona going into the game," Di Matteo told reporters.
"I think they don’t like to play against us. The results in the past have shown that - maybe the way we play is a little bit more difficult for them to play against.
"We need two perfect games against Barcelona. I think in the last six weeks we have been performing well and that given us the belief."
Di Matteo also admitted that facing Lionel Messi the current Champions League holders was the biggest test of his managerial career so far, but was bullish about his side's chances of stopping Barcelona forward Lionel Messi.
"I think for anyone facing Barcelona would be their biggest challenge," he said.
"I think it has to be a collective team effort to stop him [Messi]. But if you think about stopping just one guy, they've got nine others, so the tactic is not to stop just one player but limit their threats in general, not just Messi."
Chelsea will also be without David Luiz for the tie in London after the Brazilian picked up a hamstring injury at Wembley on Sunday, and Di Matteo bemoaned Luiz's absence.
"It’s obviously a big loss us," he said. "He is a fantastic centre half and he was in excellent form. We’re hoping to have him back within two weeks and it time for the end of the season."
Fernando Torres may be in for a return to the first XI after not starting against Spurs, and his manager praised his contribution in recent weeks and believes he should be part of Spain's squad at Euro 2012.
"He’s been doing very well especially in this last period he's been scoring and assisting goals," Di Matteo said. "His work rate has been good and I have no doubt that he should be in the team for the Euros."