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Boom mic visible
(at around 6 mins) During the science trip where Peter is bitten, Harry says that the microscope is the "largest electron microscope on the eastern seaboard". However, the microscope images shown are taken from a phase contrast microscope not an electron microscope. (Phase contrast gives less magnification but lets you work with live material, for example, the cell being manipulated in the image shown on the film).
Crew or equipment visible
Errors in geography
(at around 23 mins) When Peter is first becoming aware of his "spider sense" in school as Flash is approaching from behind, he sees a fly and a paper airplane moving in very slow motion. The fly has two wings when in reality, flies have two sets of wings, the main wings plus a smaller "halter" wings underneath (to maintain balance in flight), for a total of four wings.
(at around 1h 23 mins) When Spider-Man saves the baby from the burning building, it's amazing that the baby and the blanket are so clean - no soot, smoke damage, charring, etc. The firefighters have obvious soot around their noses from trying to breathe in that area. Did Spidey manage to give the baby a bath before presenting it to momma?
Incorrectly regarded as goofs
(at around 33 mins) When Peter is practicing his web-slinging in his room and catches the lamp to pull it towards him, it doesn't appear to move very fast off the chest and looks more likely to fall on the floor before the shot changes. When it cuts to Peter, it speeds past his shoulder a lot faster to smash on the wall behind him. Furthermore, Aunt May would have easily seen the amount of webbing all over the room through the gap in the door as she's talking to him.
Although Peter is supposed to have perfect vision after his bite, Tobey Maguire's contact lenses are visible at various points throughout the movie. It could be that Peter took to wearing "blank" contact lenses to account for the fact that he no longer needs glasses, but is more likely to be a true goof.