| 1 | 0–300m | Cavernous Cold Rock & Clay | 15.0°C | 12.6°C to 18.5°C | Fall damage, Spike traps, Geysers, Shade crawlers | Shade crawler | Geyser, Jump pad, Spike trap | Glowplant, Geotree, Hydreed, Stoneplant, Leadbush, Ceiling rye | Wooden structures, Malfunctioning Tech-Ball (Mercenary quest) | Break open the Tech-Ball to obtain and use the Nano-tech scanner before leaving Layer 1 |
| 2 | 300–600m | Pitch-Black Flooded Caves | 17.5°C | 12.5°C to 21.4°C | Total darkness, Groundwater & Lumalgae flooding, Cave tick infestations | Shade crawler, Cave tick | Geyser, Jump pad, Spike trap, Barbed fence, Bear trap, Cave tick spawner | Glowplant, Geotree, Hydreed, Stoneplant, Leadbush, Ceiling rye | Wooden structures, Concrete structures, REL-Hyper-Regeneration Booster chip site | Contains the REL-Hyper-Regeneration Booster chip site (Chip 1 of 6) |
| 3 | 600–900m | Arid Sandstone & Viscous Oil Chasms | 26.6°C | 12.4°C to 35.6°C | Viscous Oil pools (exhaustion drowning), Automated Turrets, Landmines, Tall vertical drops | Shade crawler, Sand stalker | Jump pad, Barbed fence, Bear trap, Turret, Stalactite, Landmine, Waste pipe | Sand rose, Sandvine, Cactus, Dried bush, Brownshroom, Glowplant, Stoneplant | Wooden structures, Recharging station vault (often oil-flooded), Micro-SINGLEBTR 1CHRG capacitor site | Contains the Micro-SINGLEBTR 1CHRG capacitor chip site (Chip 2 of 6) |
| 4 | 900–1200m | Radioactive Industrial Ruins | 25.8°C | -20.4°C to 38.6°C | Radiation sickness, Radwater pools, Radioactive barrels, Ceiling turrets, Extreme thermal swings | Shade crawler, Cave tick, Sand stalker | Turret, Ceiling turret, Landmine, Barbed fence, Bear trap, Radioactive barrel, Spent fuel, Sound cannon | Glowplant, Stoneplant, Leadbush, Dried bush | Wooden structures, Concrete structures, Recharging station vault, 0.2.9-Mind-Swap chip site | Contains the 0.2.9-Mind-Swap chip site (Chip 3 of 6) |
| 5 | 1200–1500m | Subterranean Jungle & Predator Den | 21.8°C | 5.6°C to 31.5°C | Elder thornback (6000 HP apex predator), Tentacle-plants, Skullcrushers, Coils | Elder thornback, Thornback, Tentacle-plant, Overgrown tick, Cave tick, Sand stalker, Shade crawler | Geyser, Jump pad, Spike trap, Bear trap, Cave tick spawner, Skullcrusher, Banana-plant, Coil | Banana-plant, Pop, Sandvine, Glowplant, Stoneplant, Ceiling rye, Geotree, Hydreed, Leadbush | Wooden structures, Brick vault, 0.0.7-Emergency Rejuvenation chip site | Contains the 0.0.7-Emergency Rejuvenation chip site (Chip 4 of 6); final layer of the Steam Demo |
| 6 | 1500–1800m | Sub-Zero Glacial Cavern | -30.6°C | -33.0°C to -24.0°C | Lethal hypothermia (-30.6°C average), Thin ice collapse, Gunmines, Snow striders, Falling icicles | Snow strider, Shade crawler | Gunmine, Turret, Spike trap, Fan trap, Geyser, Jump pad, Bear trap, Icicle, Ice stalagmite | Frigiant, Heatfern, Glowplant, Stoneplant, Leadbush | Life-pod, Ion space heater | Navigate between Life-pods and Ion space heaters to survive freezing descent to Layer 7 |
| 7 | 1800–2100m | Rain-Soaked Bioluminescent Mycelium Caves | 17.8°C | 14.2°C to 23.2°C | Constant rain causing wetness hypothermia while sleeping, Radioactive fungi, Grabber-shrooms, Sawblades | Grabber-shroom, Overgrown tick, Cave tick, Shade crawler | Sawblade, Sidestabber, Dropshroom, Bear trap, Jump pad, Cave tick spawner | Fungali bungali (exclusive source of Bunchunks for Procoagulant), Glowshroom, Brownshroom, Bouncecap, Mushroom tree | Life-pod shelters, MINI-AI chip site | Contains the MINI-AI chip site (Chip 5 of 6) |
| 8 | 2100–2400m | Distorted Crystal Abyss | 18.5°C | 16.9°C to 20.7°C | Sheer vertical drops, Volatile distorted crystals (Turbulent/Blood/Relief/Oxygen/Digestion/Soothing), Flying Manifestations | Manifestation | Crystal hazards, Geyser | None (mineral-only biome; harvest Crystal shards instead) | Crystal formations and deep vertical shafts | High-verticality traversal layer; Jetpack or grappling gear strongly recommended |
| 9 | 2400–2700m | Magma Tubes & High-Pressure Vents | 20.9°C | 10.7°C to 40.8°C | Extreme heat (up to 40.8°C), Liquid Magma pools, Barotrauma air pressure in lower half, Laser & Cryo Drones | Laser Drone, Cryo Drone | Lava Geyser, Piston, Sawblade, Fan trap, Bear trap, Stalagmite | Coldpuff, Spicevine, Stoneplant | Industrial vents, Kinetic Transmitter chip site | Contains the Kinetic Transmitter chip site (Chip 6 of 6) |
| 10 | 2700–3000m | Deepest Cargo Vault & High-Pressure Core | 22.0°C | 18.0°C to 26.0°C | Continuously rising air pressure causing lethal Barotrauma, Dense gauntlet of every mechanical trap type | None (pure mechanical trap gauntlet) | Spike trap, Barbed fence, Bear trap, Turret, Ceiling turret, Landmine, Coil, Spent fuel, Sound cannon, Skullcrusher, Sawblade, Piston, Fan trap | Stoneplant | The Cargo structure, High-powered Drill Pod (requires the Cargo to activate) | Never rolls a Layer Modifier; retrieve the Cargo here and board the high-powered Drill Pod (or loop to Layer 1 in Endless mode) |
| 11 | Ascent to Surface | Fleshy Organic Ascent Shaft | 24.0°C | 20.0°C to 28.0°C | Steep upward climb after the Drill Pod breaks down; managing accumulated trauma before reaching the Surface Lab | None | None | Crimson bush, Crimtree, Pus sack, Vein, Pop, Mushlight, Leadbush, Brownshroom, Ceiling rye, Stoneplant | Broken Drill Pod arrival point, Surface Exit (at top of layer) leading to the Lab / Surface endings | Climb to the top exit carrying the Cargo (or M.E.R.C. scarf in inventory) to trigger Bio-pod, Escape, or Mercenary ending |