Puyol has missed much of the season with knee problems but may feature as Barcelona look to bounce back from a disappointing week against rivals Real Madrid.
After being knocked out of the UEFA Champions League by Atletico Madrid, a surprise 1-0 defeat at Granada on Saturday put a dent in their hopes of retaining the Liga crown.
The return of their inspirational captain, who will leave the club at the season's end, could be the lift that Barcelona need.
Fellow defenders Gerard Pique (hip) and Marc Bartra (hamstring) have also been sidelined recently but Martino revealed on Tuesday that the diagnosis on all three was encouraging.
"We had some rather good news today regarding fitness (of the defenders) and we worked on some ideas of play," Martino said.
"We are going to make our final evaluation after tomorrow morning's training session.
"We've been watching him (Puyol), how his knee and his injury are developing. I have a very high opinion of Puyol, not just as a footballer but as a person and everything he means to the team.
"So if we do decide that he can play tomorrow, I think this could be a very important moment of the season, not just for him but for his colleagues as well."
Martino insisted that the clash versus Real has "nothing to do" with recent defeats, and rejected suggestions that winning the Copa del Rey could save their season.
He added: "This is a final. This has nothing to do with the two recent competitions. This starts and begins on Wednesday. There is no before, there is no after.
"I don't think this is seeking vengeance for what happened in the Champions League and league. It's important because it is a final.
"I don't think we should be trying to make up for things that happened in different competitions. I don't think a win in one game means that it doesn't matter that we lost before.
"I don't want to play down the importance of winning a title, neither do I want to play down the importance of what has happened in the Champions League and La Liga.
"This is a Clasico in a final and it is a very important match. But whether that would mean we feel happier about what happened in those (recent) games, no."
Martino admitted that Barcelona were "suffering" but that the absence of Real Madrid star Cristiano Ronaldo, who has knee and thigh problems, would boost his team.
He also said that if none of Puyol, Pique and Bartra were fit, they still had enough defensive options.
"At the weekend (against Granada) we realised what different options we have got for centre-back," he continued.
"The game was lost but I don't think that was because problems with defending. I think that was with problems in attack."
