Bioluminescence Lesson Documentation

The Bioluminescence lesson lets students explore an example of a bioluminescent reaction and see how temperature affects the reaction. This document covers the materials and setup for the lesson and contains general notes.


We developed this lesson using the ATP Firefly Kit from Carolina (Item # 203420). We recomend you use the larger version that's designed for a class of 30 (Item # 203421). There are similar kits from many other providers. In general, you need the following materials:

  • 8.0 pH buffer powder
  • Powdered firefly lanterns
  • ATP
  • Test tube color sensors
  • Test tubes
  • Test tube clamps
  • A hot water bath
  • An ice bath or lab refrigerator


The kit should remain refrigerated before use. Allow the components to warm to room temperature before use. The instructions in the lesson assume you will pre-mix the buffer solution and your students will be responsible for mixing the ATP solution and the firefly lantern solution as well as heating and cooling the vials. Follow the instructions in the Carolina kit to make the buffer solution.

The Carolina instructions also contain details on how the students should prepare the APT and firefly lantern solutions. The lantern solution should be transfered to test tubes that fit in the test tube color sensor before adding the ATP.


  • Some of the vials will need to be refrigerated, or placed in an ice bath for about 10 minutes. Depending on how fast your students work, they can probably place the vials in the bath before they start working on the first reaction. The lesson reminds them to do this but you may need to double check.
  • The hot water bath needs to be at least 65 C. The luciferase enzyme will quickly denature, so the vials only need to be heated for about a minute.
  • The last section of the lesson mentions a demonstration you can perform with a glowing test tube of ATP and firefly lantern solutions. Just add a few drops of sodium chloride solution to the test tube. The light should be quenched very quickly. This solution is included in both of the carolina kits.