In this level, the scientist is attacked. The scientists admits in the level's prologue that he must tighten security, but chooses to continue testing and tighten the security tomorrow. Alarms go off as the test starts, and the scientist is attacked.
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After completing the thirtieth test, the doctor injects Blue into a strange robot capsule, where the games ends with "To Be Continued".
Crawling green enzymes - Green enzymes which crawl on the ground. They can be killed when Blue shoots while crouching.
Monkey green enzymes - Green enzymes that have the ability to propel themselves up and move on the ceiling.
Mimic green enzymes - Green enzymes that change their appearance from a blob to something that looks like Blue. This transformation copies Blue's movements except for standing still. Also, when Blue fires, some time later the mimic green enzyme fires in the same place Blue did.
Shield orange enzymes - Orange enzymes which wear a shield that can protect itself from Blue's bullets. The enzyme itself can only be killed if Blue shoots the orange enzyme from its back.
Dodge orange enzymes - Orange enzymes which have the ability the dodge Blue's bullets coming from forward or behind. They can duck from bullets shot from the bottom or the top. However, it is slow from getting back up after crouching.
Flying orange enzymes- Orange enzymes which fly and convert Blue's bullets to orange bullets if one is shot to it from the front or its back. It can only be killed by being hit from its own bullets twice.
Monkey green enzymes- Green enzymes which move from the ceiling and fall when Blue when it is on top of the enzyme which can crush Blue. They can be killed when Blue manages to dodge the enzyme after it falls down and shoots it.
Sniper green enzymes- Green enzymes which uses a part of their body to throw slime at Blue and can quickly throw and regenerate themselves. Both the slime and the enzyme itself can be killed by Blue's bullets.
Bouncing green enzymes- Green enzymes which bounce around and rebound itself and travel towards another direction after hitting a wall, platform or the ceiling. They can be killed after taking a hit from Blue's bullets.
Mines - Mines explode on contact. After Blue explodes from the mine, it will automatically respawn.
Lasers - Lasers kill the player if the beam is touched. The cannon cannot be destroyed.
Proton cannons- Proton cannons shoot small bullets which can kill Blue on contact. The cannon requires two blows to be destroyed.
Detection proton cannons - Detection proton cannons hang from the ceiling. Once it targets Blue, a bullet will be shot. The cannon is destroyed with one blow.
Nitrome has said that the game went into production right after Test Subject Blue. From the trailer released on April 20, 2011, Blue is the hero and green enzymes are the enemy. A sequel is currently on development, officially known as Test Subject Complete.
April 7th, 2011
On April 7th, Nitrome released the below image in a blog post telling fans that Test Subject Green was in development.
April 20th, 2011
Nitrome released a 58 second Trailer of Test Subject Green, revealing new features, the storyline, and enemies.
May 6th, 2011
On May 6th (four days before the release), Nitrome hinted that they would be releasing a new game the next week. That game was Test Subject Green.
In level two, if Blue were to be killed by the monkey green enzyme, it would randomly start moving in between different areas between teleporters while not going through the teleporters at all. It would also become impossible for the player to destroy the enzyme or the green enzyme destroying Blue. Later, a blog post reported that the glitch had been fixed.
On level eleven, if Blue died in a certain scenario, they could be trapped with no way out. This glitch was later reported fixed.
In level fifteen, if Blue jumps on a certain teleporter right above a laser, he will sometimes die, as if he landed on the laser instead of going through the teleporter to the one above it. (See image below.)
In level twenty, on occasion Blue reaches the end and collects the food (as meant to), gets sucked up the tube (also as meant), but the level does not end and the timer continues.
Sometimes, when Blue is killed and is sent back to his nearest checkpoint, he can do a super-jump by rapidly pressing jump and shoot at the same time while invincible, which may shoot Blue off of the screen, forcing the player to restart the level.
It is possible to get to the other side of a teleporter if right and left keys are pressed steadily and fast if Blue remains in the middle of a teleporter.
If the player holds down the shoot button and the up button, right as Blue is spawning, Blue will rocket upwards in the air. Also holding down the left or right keys can adjust flight direction.
As of late June 2017, Test Subject Blue no longer advances past the startup, rendering the game's levels inaccessible.
Picture featured on the Nitrome blog with news that Test Subject Green is in development
An advertisement for Test Subject Green as it appeared in the Ad section of Nitrome.com 2.0
The logo for the game as seen on the Nitrome game slider on the front page
The link from Test Subject Blue. Interestingly, this can be seen on other sites that contain the game