Printing with GelMA: Troubleshooting
Below are some common issues users have when printing with GelMA and potential troubleshooting steps. For more details on this material, check out some background information, our protocol or our bioreport.
Issue 1: My GelMA solution seems to have a low viscosity, or quickly dispenses out of my syringe.
Troubleshooting tip 1: Try placing your syringe on ice for 10-15 minutes before loading into the Allevi.
Prior to photocrosslinking, gelatin methacrylate has a reversible thermal gelation, which causes the material to be liquid at 37 degrees celsius and a solid gel at room temperature (around 22 degrees celsius). Because of its shear-thinning properties, GelMA can be printed in this solid gel phase, offering some structural integrity prior to crosslinking.
If your GelMA appears to still be in liquid form when dispensing from the extruder (especially if it appears to gel a few minutes after extrusion), the loaded GelMA solution has likely not completely cooled to room temperature. Once loaded into the syringe, the solution can take a long time to cool to room temperature. Placing the syringe on ice prior to loading will help speed up this process.
Troubleshooting tip 2: Check your needle type, print temperature, and print pressure.
Check what needle type you are using and your print temperature and pressure settings. We suggest using a 27 gauge tapered metal Fisnar needle. Needles with a larger diameter will cause the material to extrude at a faster rate. Also, check your temperature and pressure settings. We suggest printing at room temperature (around 20-22 degrees celsius) and a pressure of around 10 psi.
When beginning a new print, it’s always best to calibrate your print pressure, first starting at around 0 or 1 psi and slowly working your way up to a pressure that causes an even extrusion. Print pressure can be affected by needle type, cell concentration and the volume of material loaded in a syringe, so it’s always best to start out with a lower pressure instead of accidentally dispensing most of your material with a higher pressure setting.
Issue 2: I have difficulty sterile-filtering my GelMA solution
Troubleshooting tip 1: Make sure your GelMA solution is protected from light.
The first thing to check is that your GelMA has not prematurely crosslinked. Does your material still appear to be gelled even after heating it to 37 degrees celsius or higher? If so, it’s possible the solution, after being exposed to light, began to permanently crosslink. Try remaking your solution, being sure to protect it from any ambient light sources.
Troubleshooting tip 2: Try heating your GelMA to 60 degrees celsius prior to filtration.
GelMA becomes slightly less viscous when heated to higher temperatures, making it easier to filter. Try heating it to 60 degrees celsius, as well as even warming your syringe and filter, prior to filtration, to make the process easier.
Troubleshooting tip 3: Try using a larger filter.
We suggest using a 0.2 micron syringe filter, but if you have significant trouble filtering the solution with this filter after trying the above steps, you can try using a larger 0.45 micron syringe filter, which will decrease the force needed to push the solution through the filter.
Issue 3: My GelMA solution keeps clogging, or has an uneven extrusion rate.
Troubleshooting tip 1: Check what needle type you are using.
We suggest all-metal tapered 27 gauge tips (about a 0.33 mm diameter) when printing with GelMA. The all-metal tip helps prevent premature crosslinking in the needle before extrusion, which could cause clogging. Likewise, the tapered shape decreasing the pressure needed to print and helps to minimize clogging of the material.
Tips with a smaller diameter than 300 microns will quickly clog when using GelMA, while tips with a larger diameter will clog less. Even with these tips however, GelMA can still occasionally clog the syringe. Readily available extra needle tips and some patience with the material are also suggested.
Troubleshooting tip 2: Add some hyaluronic acid to your solution.
Previous studies have demonstrated adding hyaluronic acid to gelatin-based formulations improves the consistency of extrusion and minimizes clogging. Depending on how much hyaluronic acid you add, you will lose some structural integrity (the ability to build in z-height without support materials), but this troubleshooting tip will help minimize clogging.
Troubleshooting tip 3: Ask about our new processed gelatin reagent.
We realize that GelMA is far from the perfect reagent for bioprinting, and we’ve been working hard at Allevi to offer you improved solutions for your research. Ask us about our new gelatin-based reagent that is specially formulated to minimize clogging and offer consistent extrusion.