The aid, he said, is scheduled for Feb. 23.
"There is much that we have to do to move and bring in the humanitarian aid," Guaido said while standing in a scaffold on the Francisco de Miranda avenue, in the same location where 17-year-old Neomar Lander died while protesting Maduro in 2017, Venezuela's El Nacional reported.
A total of 250,000 volunteers registered and will soon receive instructions on how to organize and distribute the aid.
"Those that bet that we were going to become tired have definitely lost. Venezuela is facing a war against hunger, misery and an inefficient health system," Guaido added.
"Let those that believe they can intimidate us with war exercises know they only make our Armed Forces vulnerable."
In addition to aid being collected in Cucuta, more will be gathered in Roraima, Brazil.
Guaido has said the National Assembly he leads declared Maduro's presidency illegal. As a result, assembly leadership is to take over the presidency until new elections are held. The National Assembly leader has established relations with several countries and sent representatives.
Guaido has organized two other protests so far this year in Venezuela calling for the immediate transfer of power from Maduro while a free election process is held.
Maduro has rejected calls for new elections.
"What's the logic, reasoning, to repeat an election?" he asked in an interview with BBC News.
Maduro also accused the United States of "warmongering" to take over Venezuela.
Guaido said last month he will lead the transition but insisted rebuilding Venezuela democratically must begin with the delivery of humanitarian aid to save hundreds of thousands in peril.
Maduro has maintained a grip on power and rejected the aid, saying it's a small fraction of what his government loses because of what he considers unfriendly U.S. policies. Not only has the U.S. government sanctioned Venezuelan gold and oil exports, but officials close to Maduro have been the target of criminal investigations.
The divisions in Venezuela between Guaido, a 35-year old engineer who was little known just weeks ago, and Maduro, who is supported by the military, has extended into a global split.
Some nations including the United States, most Latin American bigger nations and most European and Baltic nations recognize Guaido as Venezuela's leader -- while others like China, Russia, Cuba and North Korea do not.
Maduro denies Venezuela is undergoing a crisis and there have been marches supporting his claim that the situation stems from U.S. efforts at organizing a coup.
More than three million Venezuelans, or about a third of the population, have left the country in recent years to escape political violence, hyperinflation and lack of medicine and food.
Leaders like Mexican President Andres Manuel Lopez Obrador have been critical of the U.S., Canada and other nations' decisions to coordinate joint assistance at a time when such actions can be easily confused with politics.
Lopez Obrador said he favors assistance, but it should be provided to other at-risk populations like those in Haiti and areas of Mexico. He said it would be better for the United Nations, instead of politicians, to handle the aid.