Tear Jerker / The Spectacular Spider-Man

• The look of defeat on Peter's face when the symbiote asks him to join with them. After having one of his worst memories dug up, he looks like he's crossed the Despair Event Horizon as he starts to accept. Thankfully the memory of his uncle steps in to snap him out of it.
• A lot of episodes make it clear that Peter often outright hates being Spider-Man, and it's never more apparent than during the flashback sequence in "Intervention". When the symbiote forces Peter to relive his origin, the first thing he tries to do is change what happened, reasoning that if he isn't bitten by the spider, Ben won't die. His thoughts are interrupted when the spider bites him, causing him to let out an anguished scream.
  Peter: Not again... NOT AGAIN!!!
• Peter's reaction when he finally has the iconic moment of recognizing the burglar's face. At first he's silent, as his earlier remark of "Not my problem" repeats in his head over and over. Then, it shows Ben with a look of horror on his face, flinching alongside a sound reminiscent of a gunshot. Finally, Peter whispers to himself in horror.
  Peter: What have I done?
  • Josh Keaton's performance really sells the scene. He makes Peter sound moments away from breaking down and sobbing.
• Eddie Brock's final refusal of friendship from Peter. Going so far as to say how he really feels about him.
  Venom: WE'RE NOT BROTHERS!!! Our parents may have died together, but you had your precious Aunt and Uncle. We had no one. We've always been alone... until now.
  • What makes it worse is that this isn't like most versions of their rivalry. Just like in the Ultimate comics, these are two best friends who clearly needed to be there for each other and yet due to miscommunication, and because of the symbiote, the two started to become more distant and Eddie slowly became one of the most dangerous enemies Peter would ever face. It just goes to prove the old saying: 'There is no worse enemy than an old friend.'
  • Peter's response is both badass and heartbreaking as he's hurt by how Eddie truly feels about him, is sickened by the petty motivation for his villainy, and for endangering everyone he cares for.
  Spider-Man: Then we're DONE talking.
• When Peter Parker broke up with Liz Allan, she created a huge scene specifically meant to gain the attention of the crowd, making it look like she was doing the dumping instead. As it turns out, this was actually a way of hiding her pain, because immediately after she "storms off", she is seen hiding behind a corner, crying her eyes out. This version of Liz Allan actually had genuine feelings for Peter. It really shows the strength of the writers when they can pull off emotional scenes even with relatively minor characters from the comics.
  • The way he does it is pretty sad too, as he admits that she's been a really great girlfriend, but that he feels that they'd be better off as friends.
  • Worth noting that this breakup from Peter comes after Liz slowly builds interest in him over the course of two seasons, and followed by Peter frequently blowing her off due to his secret duties as Spider-Man, clearly beginning to favor Gwen over her, standing her up on the night she had a play recital her parents were attending. The fact that it takes place shortly after her brother gets arrested for supervillainy is just an added layer of pain for a character already going through so much.
• Norman Osborn's callous treatment of Doctor Octavius is also this. Otto is a timid man who Norman browbeats into performing extremely unethical experiments, verbally abuses and calls "weak" and eventually tries to murder to keep his secrets. The sight of Otto sobbing and begging for help is heartbreaking. And the first thing he does after his rebirth as Doctor Octopus? Lash out at Norman for all the abuse he took.
• The ending of "The Uncertainty Principle". Harry is revealed (falsely) to be the Green Goblin and has been drinking the addictive and unstable Globulin Green. Norman is pissed and demands answers. Harry throws him at a wall and tells him that he only wanted to be the son he wanted. He then keeps switching moods due the prolonged addiction. It was both sad and scary.
  • And we later find out that said father he was ranting at had just got done setting him up as the Green Goblin, which included twisting his leg. Which makes this scene even sadder in hindsight.
• The fate of John Jameson/Colonel Jupiter. Knowing that this is the same person we saw as a normal man who saved a building from the Green Goblin makes it even worse to see him as a crazed psycho.
  "I miss the power... I need the power... GIVE ME BACK THE POWER!! GIVE IT BACK!!"
  • Made even sadder when you realize who's in the cell next to him, and how their situations are VERY similar. And it's one of the few times we see J.J. showing anything other than anger or smarmy condescension. The look on his face is heartbreaking.
  • An extra layer of hurt? When he and Spider-Man worked to save civilians from the fire, it's clear in Spidey's voice that he's happy to have a second superhero in New York, that maybe it doesn't all have to fall on him anymore. Of course, then the rest of the episode happens.
  • The fate of Mark Allen, aka Molten Man is very similar.
• When Eddie is separated from the symbiote again in "Identity Crisis" he breaks down, crying out for it to not leave him again and even begging Peter help him get it back when it escapes down a drain. He's ultimately carted off bound to a stretcher, frantically screaming that the symbiote will come back. The dependency on the symbiote he's developed really makes you feel sorry for the guy.
  Spider-Man: Eddie! Eddie, it's over. You're free, you can stop this. Stop... hating.
  Eddie: (on the verge of tears) Can't... It only loves me... f-for the hate...
• Electro in general was an emotional character. In his early appearances, he didn't want to be what he was. It was just a cruel twist of fate. His "condition", however, slowly consumed his mind and drove him a little nutty- though that doesn't really make him slightly less sympathetic.
  • The earliest examples are his talking to Eddie about how he had plans to go back to college, meet someone and have a life, all taken by the experiment. When he tries to get out of isolation by having a coffee, people stare at his containment suit and he finds he can't even drink or eat anything, leading to him unleashing his powers and shorting the electrical gadgets. He calls out that he's sorry before running away. He sounds absolutely heartbroken. Of course, shortly after, Spider-Man attacks him (believing him to be a crook running from a crime scene) pushing him down the path of villainy.
• Harry watching Spider-Man accidentally "killing" the Green Goblin, his father, in the series finale. Peter's expression of guilt and subtle despair reflect his failure despite everything he could do (which is very similar to what happened to his uncle).
  • The whole episode could be classified as a giant tearjerker. Let's recap: Dr. Connors and his wife are being blackmailed to quit their jobs, Peter's best friend Harry has been manipulated into thinking he's the Goblin (and all it supposed for him) only to discover it was all a lie, he's lost his father, Peter ends up mostly alone as his relationship with Liz ended on a really painful note, he's unable to be with Gwen (after finally revealing their feelings for each other) because she can't break up with Harry as planned because he's grieving (it's hinted that he's consciously using pity to keep Gwen with him because he heard the aforementioned reveal), and his friendship with Harry is likely strained as the latter now hates Spider-Man for killing his father and learning of Peter and Gwen's feelings for each other (resulting in the above mentioned manipulation). Many people will consider Spider–Man a criminal for what he's done and even when Peter monologues about New York being safe for now on, we know it won't last long, especially because we see Norman is still alive and will surely return to cause more problems. And to top it all off, since the show was cancelled, it will never have a satisfactory conclusion. Downer Ending at its finest.
  • It gets worse: in Spider-Man: Across the Spider-Verse, George Stacy is revealed to have died somewhere down the line, not only undoubtedly causing Gwen further heartbreak but adding onto Peter's turmoil as well. This Spidey appears in the film, but it's shown that he Took a Level in Cynic. Not only did his George Stacy (the only adult who knew about his secret identity and helped him how/whenever he could) die tragically, but Josh Keaton confirmed that this Peter Parker's life began to spiral downward after the series finale: George Stacy died, Harry and Gwen lost faith in Spider-Man (putting a serious strain on their friendship with Peter), and his mental health took so much of a toll that he agreed to join O'Hara's Spider Society (possibly leaving his own dimension in the process, much like Spider-Woman Gwen Stacy).
• Spidey working with Uncle Ben's killer, Walter Hardy (the father of Felicia/Black Cat). Every word out of his mouth is rushed and aggressive, giving the sense he's only barely keeping himself from falling apart in the man's presence. Even at the end, when the man chooses to sacrifice his chance at freedom to contain the criminals Peter can't ever bring himself to change his emotions.
  Spider-Man: That won't make us even... you're not forgiven!"
  Walter: (sadly) I understand.
  • Black Cat won't forgive him either, for causing her father to stay behind, which leaves Spidey somewhat guilty, but likely still angry at her father. When Felicia is first driving away, she has angry tears streaming down her face. She comes back and saves Spidey from Green Goblin, but cuts off his thanks and yells that she'll never forgive him for what he did to her father, losing himself another ally in the process.