Videotape of the moment Steve Irwin was hit by a stingray's tail shows the Australian naturalist pulling the barb from his chest, his manager has said.
"The tail came up, and spiked him here [in the chest], and he pulled it out and the next minute, he's gone," Mr Irwin's manager, John Stainton, said.
Queensland state police have now taken the tape to be used in an inquest into the incident on the Great Barrier Reef.
The much-loved TV star could be given a state funeral if his family agree.
Throughout Monday and Tuesday thousands of fans gathered at Mr Irwin's zoo in Beerwah on Australia's sunshine coast to lay flowers and write messages of condolence.
On Tuesday, Australia's federal parliament paused to honour Mr Irwin, whom Prime Minister John Howard said had died in "quintessentially Australian circumstances".
Mr Howard quoted Australian-based actor Russell Crowe, who had paid tribute to Mr Irwin earlier by saying: "Steve Irwin was the Australian many of us aspire to be."
Mr Irwin had been in the water at Batt Reef, off the resort town of Port Douglas about 100km (62 miles) north of Cairns, filming bull stingrays for a TV documentary called Ocean's Deadliest.
'It probably felt threatened'
Cameraman Ben Cropp, who was also on the reef when Mr Irwin was killed on Monday, spoke to a member of the production crew who had seen the footage of the incident.
"He was up in the shallow water, probably 1.5m to 2m deep, following a bull ray which was about a metre across the body - probably weighing about 100kg, and it had quite a large spine," Mr Cropp told The Australian newspaper.
"It stopped and went into a defensive mode and swung its tail with the spike. It probably felt threatened because Steve was alongside and there was the cameraman ahead, and it felt there was danger and it baulked."
Members of the Dasyatidae family of cartilaginous fish, with about 70 species worldwide
Mostly found in tropical seas, but exist in freshwater too
Feed primarily on molluscs and crustaceans on sea floor
Swim with flying motion using large pectoral wings
Usually docile, not known to attack aggressively
Equipped with venom-coated razor-sharp barbed or serrated tail, up to 20cm long
Though armed with a venom-coated, sharp barb on their tail, stingrays only use the weapon defensively and attacks on humans are extremely rare.
Appearing on ABC News, Mr Stainton, who was with the TV crew on the reef, described watching the footage of the incident as a "terrible" experience.
"It was a hard thing to watch because you are actually watching a person die," Mr Stainton said.
Mr Irwin's heart is believed to have been pierced, and he died almost immediately.
Mr Irwin's body has now been flown from the mortuary in Cairns, where a post-mortem examination was carried out, to the small town of Beerwah where he lived.
The TV presenter is survived by his wife Terri and two children - eight-year-old daughter Bindi Sue and three-year-old son Bob.
No funeral arrangements have been announced yet, but Queensland State Premier Peter Beattie said Mr Irwin would be given a state funeral if his family approved.
Mr Irwin was regarded by many Australians as a national treasure and as news of his death broke, news websites across the country found it difficult to cope with the demand for information.
Tributes flowed in from Mr Irwin's fellow conservationists and his many fans around the world, and across the country on Tuesday the morning papers devoted their front pages to news of his death.