A smiling Guaido could be seen in midmorning entering the Federal Palace building that houses the National Assembly -- the only government branch under opposition control -- alongside other lawmakers on a video shared by his press team.
"Yesterday the dictatorship tried to prevent our session but they couldn't and they can't," Guaido wrote on twitter.
"Today we will sit in session honoring once more the support and confidence of the whole of Venezuela."
SEBIN security agents and the National Guard, which provides security for the building, had prevented lawmakers from entering on Tuesday.
It was the latest move in a series of measures taken by the Maduro regime against opposition lawmakers since Guaido's failed April 30 uprising.
Venezuela has been in political turmoil since parliament speaker Guaido declared himself acting president in January in a direct challenge to Maduro's authority.
The Constituent Assembly, set up by Maduro to replace the sidelined National Assembly, has stripped a dozen opposition lawmakers of their parliamentary immunity.
The Supreme Court has likewise charged 14 deputies with involvement in the failed uprising when Guaido was joined by around 30 members of the armed forces in a revolt that quickly fizzled out. It did however spark two days of deadly clashes between protesters and security forces.
The military largely remained loyal to Maduro, though, strengthening his position in the power struggle with Guaido, who is recognized as interim president by more than 50 countries.
Of the lawmakers singled out by authorities, deputy assembly speaker Edgar Zambrano was arrested by SEBIN agents last week. Another fled to neighboring Colombia while four others sought refuge in diplomatic compounds.
Maduro's right-hand man Diosdado Cabello justified Tuesday's intervention by claiming there was a bomb threat in the building.
Guaido accused the security forces of using "brute force" and said the building was "occupied militarily."
Tuesday's session was meant to discuss government measures taken against National Assembly lawmakers, which was instead on Wednesday's program.
"We're already here inside. Entry was as normal, they only asked deputies for their identification," lawmaker Arnoldo Benitez told AFP.
While deputies were allowed in, the National Guard prevented the press from entering, something that has happened before.
"They didn't allow in the press and that's a concern for us because the media is a shield for us here inside," Benitez said by telephone.
A line of uniformed officers carrying shields were stationed outside parliament's entrance.