Lifeink® 200 and Riboflavin Crosslinking

After printing acellular constructs with Lifeink® 200, the collagen constructs can be thermally crosslinked at 37°C.


For additional crosslinking, the acellular structures can be exposed to riboflavin and UV light. This additional crosslinking can approximately double the gelled stiffness. This protocol is not optimized for cellular bioprinting.

For crosslinking Lifeink® 200  constructs with encapsulated cells, we recommend mixing the bioink with Lifeink® 100, Lifeink® 300 or Lifeink® 400

Lifeink® 200 and Riboflavin Crosslinking Instructions

  1. Bioprint an Acellular construct using Lifeink® 200

  2. ​Mix riboflavin at a 0.02% solution (20 mg/100 ml) in 1X PBS.

  3. Bathe your construct in the riboflavin solution for 5 minutes. 

  4. Expose to UV light (365nm) for 5 minutes.

  5. Wash out the riboflavin with cell culture media (ensure the riboflavin is fully exchanged out for media).

The resulting collagen construct should have an ~2X increase in gel stiffness.

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