ATP Quantification

Introduction

Follow these step-by-step instructions to perform an ATP quantification for 3D bioprinted constructs. This protocol uses a luminescent assay to determine the number of viable cells in culture.

Materials

• CellTiterGlo 3D reagent 
• Cell culture medium (stored in refrigerator)
• Pipettes and Pipette tips
• Black 96 well plates
• At least 5 samples of all groups to be tested

Methods for ATP Quantification

1. Remove CellTiterGlo 3D solution from freezer and allow to thaw to room temperature on counter. 
   1. Note: Do not thaw this solution in water bath – container may break. Mix with media at a 1:1 ratio.
2. Add mixture of media and Celltiterglo to samples (for samples in 24 well plate, add 600 ul). Allow to incubate for 1 hour and 20 min at 37 C on a shaker. You will need at least 5 samples per group. Be sure to include negative control wells.
3. After incubation, take 200 ul of solution from each sample and pipette into a black 96 well plate.
4. Record luminescence following the CellTiterGlo protocol on microplate software.
5. To quantify data, save each group in an excel sheet in the format below and follow matlab code detailed in matlab entry.

We hope this protocol helps you easily determine the number of viable cells in your 3D bioprinted construct using an ATP quantification method. For more assay protocols – click here.