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- The test tube is covered in black. Some believe this is a reference to Touhou 6, where the stage 1 boss, Rumia, could control darkness.
- A piece of poo from Nintendo Land's Pikmin adventure. A joke on the "number 2" joke.
- a coin from Mario.
- A sign saying "4=evil". This is a reference to the fact that the word for 'four' and 'evil' are the same in japanese.
- A primid.
- labelled '6', but with paint saying "mane" above it. nothing is inside. The label is a reference to My Little Pony's six main characters, the "mane 6".
- A muffin. If the player stand infront of test-tube 6 or 8 for 20 seconds, Derpy Hooves from My Little Pony will appear out of the same portal Rainbow Dash used, take the muffin, wink to the camera and goes back throught the portal. (The number may be a reference to the fact that Derpy is used as LuckyEmile's logo. The Lucky number is 7.)
- Contains the Game and Watch Octopus. A reference to an octopus having eight legs.
- labelled ⑨, Cirno appears in the test tube. If the player stand infront of it, the music is replace with Cirno's theme: Beloved Tomboy Daughter.
- Captain Basalix from Super Mario Bros. Z. It is a reference to how LuckyEmile almost quit near finishing the project, which the creator of SMBZ, Alvin Earthworm, did. Since the tenth tube is the last one, showing the end.