Since Nuzlocke revolves around the concept of ''VideoGame/{{Pokemon}}'' getting KilledOffForReal, it's only natural that it would be filled with these moments.

!!!Main series:
[[caption-width-right:350:"Our suffering had only begun."]]

* The pictured example. It has Ruby's first death on the team in the form of a poisoned Beautifly. What hurts more is that [[HopeSpot he was just a few moments away from the Pokemon Center]].
** Repeated and arguably even sadder with Whiskers the Rattata, the first casualty of the [=FireRed=] run; the HopeSpot actually shows him getting healed before revealing it was all in the mouse's head [[GoOutWithASmile as he dies happy]].
* The finale of Season 1. [[spoiler: Steven is down to his last Pokemon, Metagross. Ruby commands Absol to use Perish Song, as he knows that none of his remaining Pokemon can take Metagross one on one. On that same turn Absol is KO'd by a Meteor Mash, 3 Perish Song counters remaining. Ruby follows with Katie, and she is easily disposed by an Earthquake from Metagross, 2 Perish Song counters remaining. Now Ruby is down to his last Pokemon, Sceptile, and as a HopeSpot arrives he is reminded of all the Pokemon he lost to get there, and that everything was for a reason; he just had to survive until the end of the turn for the Perish Song to take effect.]]
** Then [[spoiler: Metagross used Hyper Beam. And everything faded to white.]]
--> [[spoiler:'''Nuzleaf:''' "And that ends the story of Ruby, a trainer who believed he had a great destiny, that everything was happening for a reason. Do you know what his last thoughts were, as he watched his first Pokemon die? "I just don't understand." [[LampshadeHanging Now isn't that just the saddest thing you ever heard?]]"]]
* [[spoiler:Gulliver's]] HeroicSacrifice.
* [[spoiler:Pony's death definitely qualifies.]]
* Episode 29. [[spoiler: Buddy, Doakes, Blamo. You shall be missed.]]
** [[spoiler: Buddy's line: "Right... just one turn." as he [[GoOutWithASmile grins]] is particularly heart-wrenching.]]
* Season 2, Episode 19 is probably the most heartbreaking non-death strip in the whole series. [[spoiler:It details the end of Charmander and Squirtle's friendship, caused by Ruby's challenge.]]
* [[ "You have to remind it what's important."]]
* The [[ What is the Nuzlocke Challenge?]] page.
* And then during the showdown with Mewtwo, [[spoiler:Sawyer is killed and Bruce is separated from Ruby, leaving him with no Pokemon left]].
* Blastoise returning to Pallet to visit Bulbasaur, and both of them sitting under the stars in memory of Bruce.
* Ruby gets his first casualty in ''White: Hard Mode'' with [[spoiler: Hank dying to N's Pidove]]. What makes it worse is that it was displayed in the form of a GoryDiscretionShot.
* After finally beating an opponent, [[spoiler: and avenging Hank, Senor Fluf is the next to go down. [[GoOutWithASmile His last words to Ruby ask if he saw him beat N's Pidove, proud of his accomplishment]], right before he closes his eyes for good.]]
** Those same tears become bitter [[spoiler: when N walks up and hits Ruby with a simple truth, that in turn potentially reopens every single tearjerker death in the past. "Yes, it is sad... But remember... They wouldn't have died... if you hadn't caught them."]]
* The reveal of [[spoiler:Isis']] identity. Even Katelyn is disturbed by what she sees...
--> Katelyn: "...holy [[CurseCutShort fu--]]"
* Meta-example. On late Feb 2017, the author would had his car [[ broken into]] and have his 3DS and ''all'' his Pokémon games stolen. This meant that that he lost all of his friends in real life whether they lived through the story or not. The only legacy left of them would be the webcomic itself where he [[ already written down]] the notes for White.

!!!Emerald Comic - A Shoeless Adventure:
* Chapters 10 and 11. Each Pokémon gets a well-developed character, and when they die, it hits hard.

!!!Freddy's Nuzlocke Adventures:
* Towards the end of his first run comes [[ this]]. The music ''does not help''.

!!!Landwalker's Nuzlocke:
* [[spoiler: Rikishi's]] HeroicSacrifice during the [[spoiler: Champion battle.]] Complete with a flashback.
* In Landwalker's Yellow run, [[spoiler: the death of Blue's Raticate, trampled by evacuees when the SS Anne burns/sinks. Complete with shots of Blue desperately screaming and searching for it.]]
** Also, [[spoiler:Lun the Diglett's death.]]
** [[spoiler:Amy's death to a self-destructing Koffing. Red's reaction after all of it concludes ''does not help''.]]

!!![=NyaChan's=] Nuzlocke:
* In Pearl, she loses [[spoiler: her starter, who's been through ''everything'' with her]] to [[spoiler: ''[[PowerfulButInaccurate Fissure]]'', [[UndignifiedDeath of all things]], during the Elite Four.]]
* The end of the Pearl run. [[spoiler: DownerEnding doesn't even ''begin'' to cover it...]]

!!!Kynim's Diamond Nuzlocke:
* Page [[ 21]] and [[ 22]]. [[spoiler: Josie the geodude dies from poison]] The "no heal items" rule bites back.
* [[ Fantina Gym Battle]] with [[ sad death video]] [[spoiler: Apparently ghost Pokemon can die]]
* [[ Flint Battle]] with [[ another sad death video]].

!!!wizard898's Moemon: Hard Mode:
* A heartwarming picnic takes on a bittersweet air when Saya goes to help clean off her master's mouth and he rebuffs her.
* [[spoiler: Ruru's]] HeroicSacrifice, especially as her [[spoiler: Static kicking in]] proves key to their victory. The expression [[spoiler: poor Chu]] makes as it sinks in just ''makes it worse''.
* Chapter 21: [[spoiler: Volt's death. Just to see a poor sweet little girl implied to be beaten to death...that just sucks. Doubles as a YouBastard moment for Team Aqua.]]
* 22 rubs salt in the wound, as Tooh blames their trainer for what happened, calling him weak and refusing to follow him any longer. [[spoiler: Umpa joins her.]]

!!!Nonparael's Nuzlocke Runs:
* The three-part Episode 10 of the Moemon LG run, combines this with WhamEpisode: first, Gale Oak reveals he ''knows'' what the heroine is trying to accomplish, calling her cause 'crazy'. Then [[spoiler: he forces Sienna's charmander friend Allie to kill Sienna, followed up by Tia's death. The heroine reveals herself as a moemon Weepingbell, but Gale ''has her pokeball'', and Nail performs a HeroicSacrifice]]. To rub it all in, Gale [[EvilGloating mocks her afterwards]].
* In the Black run, [[spoiler: Nineveh's death]] from [[ Part 43.]]

!!!Bruce Wayne's Platinum Nuzlocke:
* The death of [[spoiler: Barbara]] against [[spoiler: Jupiter's Skuntank]]. The reactions from the rest of the team only serve to make it all the more tragic and heartrending.

!!!Mike's Crystal Nuzlocke:
* [[ Shuckie's death]] is quite saddening. It's amazing to see the death of a Shuckle being so sad.

!!!Lenni's White Run:
* It's like [[spoiler: watching your dog die]]. [[ No, literally.]] To make matters worse [[spoiler: his body shows up in later pages during the rest of the battle.]]
** [[ Also, this.]]

!!! Aouli's "The Meek" LG:
* The death of [[spoiler: Kat the Raticate]] in Rock Tunnel. The author's description of her own reaction as it happened qualifies too.

!!! Robin's Emerald Nuzlocke:
* After spending about four pages rounding out [[spoiler: Lassie's]] character [[spoiler: he's killed by a random kid's Nincada.]]

!!! "It's a Hard Life" Leaf Green Nuzlocke:
* When half the cops in Kanto are after Robin and trainers keep trying to kill her for the reward, Lt. Surge is the first person who actually tries to help, treating her as a child instead of a criminal. But the combination of bad luck, miscommunication, and a terrified, homeless and ''armed'' 10-year-old means [[spoiler:Elektra loses her remaining eye.]]

!!! Nix's Fire Red Nuzlocke:
* Rick [[spoiler:being killed by a wild Spearow. Nix is devastated.]]
** Said wild Spearow is caught and named Dredd. He evolves into a Fearow, [[spoiler:only to be killed by Misty's Starmie]].
* After being told that she couldn't control her Pokemon in battle, Nix orders Charlie to "go wild," and this leads to Charlie [[spoiler:burning Rupert (Gary's Raticate) alive. Surprisingly, this is treated as a GoryDiscretionShot.]]

!!!Jet's Black Nuzlocke:
* Our first casualty: [[spoiler:Broccoli the Pansage]], dying to save the Munna he has a crush on.
* [[spoiler:Bottles']] death, not helped by him being a Kamina expy.
* [[spoiler:Lilly]]'s death may be the worst so far--[[spoiler:we get a five second HopeSpot before she narrates a heartwrenching monologue about how unfair it is that she has to die now and how much she wishes it could have lasted longer as she plummets to her death. Her funeral afterward is also heartbreaking, with Jet wallowing in guilt and her Pokemon crying at Lilly's grave. Special mention goes to Firedog, who just sits there at her grave, [[HeroicBSOD too distraught to even say anything]].]]
* Jet getting pushed so far into the DespiarEventHorizon that after the fourth gym she honestly tries to ''join N and Team Plasma'' and is only prevented from doing so by Juniper showing up.

* [[spoiler: Smee's sacrifice.]] Even worse was that this was the first death, causing Rooke to [[HeroicBSOD want to quit]] in fear of killing any more of her Pokemon.
* Lucy revealing her feelings for Donovan, only to get rejected.
* Many of the characters' backstories are pretty sad:
** Kindle definitely has the saddest backstory. His abusive trainer refused to pull his best friend from battle, causing him to die. Angry, Kindle ran away from his trainer planning to never get caught again. He was.
** Miss was constantly ridiculed for being a female Machop, causing her to fer evolving. In Chapter 9, when a ray causes every Pokemon to evolve, Miss is forced to suffer to prevent herself from doing the same.
* [[spoiler: Oji's]] death was also pretty sad.
* [[spoiler: Granite's]] death was one of the harsher ones, especially seeing how far he has come since the beginning of the comic.
* Chapter 8 was an emotional rollercoaster:
** [[spoiler: Mystery's death is by far the saddest moment in the comic, especially given how unexpected it was. Even more heartwrenching was the characters' reactions; Scout breaks down into tears and goes into HeroicBSOD, Miss takes her anger out on everything, and Granite begins to sob into Mystery's burned tophat.]]
** And if that's not enough, only a few pages after, [[spoiler: Sam almost dies, causing Kindle to break down for the first time in the entire comic. However, she survives, and evolves into an Espeon.]]
* The ending is incredibly sad as well. [[spoiler: Rooke ultimately decides to leave, and says goodbye to all of her friends. However, right before she arrives home, she decides to turn back, but it fails. She spends the next six months trapped at home, thinking that she can never return to the Pokemon world, before seeing Lugia again.]] This ending rivals the Pokemon Mystery Dungeon endings in terms of sadness.