Petr Jiracek's well-taken goal rounded off a clinical 72nd-minute counter-attack by the Czechs to secure the win they needed to progress to the knockout phase of a European Championship for the third time in their history.
That was not before high drama in the dying seconds, when Michal Kadlec blocked a goal-bound shot just moments before the final whistle to ensure his team became the first to top their group with a negative goal difference.
Poland become only the second host nation, after Austria four years ago, to be eliminated at the group stage as they failed to claim their first ever win in the competition.
The Czechs' victory also eliminates Russia, who lost 1-0 to Greece in the other Group A clash happening simultaneously in Warsaw.
At the kick-off on a rain-sodden night in Wroclaw, both sides knew that a win would definitely send them through to the quarter-finals, but Poland did not have the luxury of a draw possibly being enough for them.
Poland manager Franciszek Smuda retained the services of Przemslaw Tyton in goal despite the fact that Wojciech Szczesny was back from the suspension incurred by his red card in the opening draw with Greece.
The co-hosts began the match the brighter in front of their home support, even if the colours favoured by both nations meant all four corners of the ground were red-and-white.
The Poles registered six shots in a dominant early spell, with two early efforts getting the home supporters off their seats as they hit the side netting. Robert Lewandowski fired wide on 11 minutes as he sought to capitalise on a sloppy backpass in the Czech defence, while defender Sebastian Boenisch's long-range strike forced Petr Cech into action with a low save tipped around the post.
The Czechs gradually began to work their way back into the match as the high early tempo brought on by the conditions and the occasion subsided. Petr Jiracek almost picked out Milan Baros's clever run with a ball dinked over the defence, but the Galatasaray striker could not quite reach it after beating the Polish offside trap. Jaroslav Plasil's deflected shot was no problem for Tyton, whilst Vaclav Pilar started to torment Poland on the left wing but was also unable to really test the keeper as the first half ended goalless.
Just after the half-time whistle blew in Wroclaw, Giorgos Karagounis's goal against Russia in Warsaw changed the complexion of the group completely, as it meant a draw was no longer sufficient for the Czechs either.
However, it was the Czechs who took the initiative armed with the knowledge that they too needed a win. They won several free-kicks around the edge of the Polish box, most of which were defended well but one came through to Tomas Sivok. The defender sent his header down into the turf, but it rebounded off Tyton's legs as it bounced back up and towards goal.
The Czechs finally made the breakthrough as the match entered the final 20 minutes. Rafal Murawski lost possession in the opposition half and they broke at pace. Baros surged into the Polish box and laid the ball off to Jiracek. The Wolfsburg attacker cut inside his marker and sidefooted a low finish underneath Tyton and into the far corner.
Tensions ran high as the clock ticked down as a scuffle broke out due to Poland's players being unhappy with the length of time Pilar was taking to leave the field as he was substituted. Plasil and Jakub Blaszczykowski were both booked by Scottish referee Craig Thomson for their part in the fracas.
Blaszczykowski also got a measure of revenge in the last few second of the four minutes of added time when his strike from the edge of the box was bound for the top corner. However, Kadlec leapt in to glance it off target with his head, and Thomson blew the final whistle just seconds later.
The Czechs had to wait for one agonising minute before confirmation arrived that Greece had won to finish as runners-up and send both of them through.
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